pp 1075-1082 PERSPECTIVES
George Shull’s 1908 seminal article ‘The composition of a field of maize’ marked the ‘exploitation of heterosis in plant breeding, surely one of genetics’ greatest triumphs’. Hybrid corn became a ‘symbol of American agriculture’ and ‘the paradigm for all developments of F1 hybrid crop varieties and more generally breeding. But there is still no consensus on the definition of heterosis while its biological basis, causal factors and genetic mechanisms remain ‘unknown’, or at best ‘poorly understood’. It is thus logical to reverse the usual approach from the exploitation of a mysterious heterosis to the triumph of hybrid corn and focus on what breeders and geneticists do rather than on the theoretical reasons for their success. This factual approach produces surprising results: (i) hybrid corn extends the isolation technique of autogamous cereals to the allogamous maize; (ii) a ‘hybrid’ is an ordinary corn plantmade reproducible by the breeder and only the breeder. It is proprietary rather than ‘hybrid’; (iii) for all practical purposes, heterosis is irrelevant; (iv) Shull justified his ‘hybrid’ breeding method by the ad hoc argument of maize ‘hybrid vigour’ which in 1914, he conflated under the name of heterosis with Edward East’s concept of physiological stimulation due to heterozygosity; (v) hybrid corn can increase yield only once and by a small margin and (vi) the huge yield gains of the last 80 years came from mass selection, a process inconsistent with the theory of heterosis. In conclusion, the enduring success of ‘hybrid’ corn was achieved at the expense of farmers, common welfare and biodiversity and dovetails with the industrial agriculture requirements of crop uniformity and breeder monopoly over reproduction. This critical understanding of the paradigm of plant breeding could have important implications for breeders and geneticists.
pp 1083-1095 RESEARCH ARTICLE
Pentatricopeptide repeat (PPR) gene family plays an essential role in the regulation of plant growth and organelle gene expression. Some PPR genes are related to fertility restoration in plant, but there is no detailed information in Gossypium. In the present study, we identified 482 and 433 PPR homologues in Gossypium raimondii (D5) and G. arboreum (A2) genomes, respectively. Most PPR homologues showed an even distribution on the whole chromosomes. Given an evolutionary analysis to PPR genes from G. raimondii (D5), G. arboreum (A2) and G. hirsutum genomes, eight PPR genes were clustered together with restoring genes of other species. Most cotton PPR genes were qualified with no intron, high proportion of α-helix and classical tertiary structure of PPR protein. Based on bioinformatics analyses, eight PPR genes were targeted in mitochondrion, encoding typical P subfamily protein with protein binding activity and organelle RNA metabolism in function. Further verified by RNA-seq and quantitative real-time PCR (qRT-PCR) analyses, two PPR candidate genes, Gorai.005G0470 (D5) and Cotton_A_08373 (A2), were upregulated in fertileline than sterile line. These results reveal new insights into PPR gene evolution in Gossypium.
pp 1097-1105 RESEARCH ARTICLE
Knowledge and analysis of the genetic structure of an endangered species is important for its conservation and evolutionary process. Simple sequence repeats (SSRs) and amplified fragment length polymorphisms (AFLPs) were used in evaluation of the genetic diversity and population differentiation in Limonium bicolor (Plumbaginaceae), an endangered herb with high medicinal and horticulture value. A total of 117 alleles were detected with an average 5.85 alleles per locus using SSR and 222 bands from AFLP were amplified in six populations. It was found that L. bicolor was characterized by high levels of genetic polymorphism (100 and 83.78%), low levels of total genetic diversity (Ht = 0.2824 and 0.2424), and moderate degrees of genetic differentiation among populations (ΦST = 0.284 and 0.251). Analysis of molecular variance (AMOVA) revealed that the main variation component existed within populations (71.56%; 74.93%) rather than among populations (28.44%; 25.07%). Four main clusters were displayed in the UPGMA using TFPGA, which was consistent with the result of principal coordinate analysis (PCA) using NTSYS. Mutations or infrequent gene flow among populations can increase the plant slowly, thus in situ conservation policies should be implemented first for effective and sustainable development. At the same time, ex situ measures, such as those individuals with rare alleles, to maintain the relationships between individuals and populations are also proposed.
pp 1107-1117 RESEARCH ARTICLE
To investigate the role of miR-27b in sheep skeletal muscle development, here we first cloned the sequence of sheep pre-miR-27b, then further investigated its expression pattern in sheep skeletal muscle in vivo, the relationship of miR-27b expression and sheep skeletal muscle satellite cell proliferation and differentiation in vitro, and then finally confirmed its target gene during this development process. MiR-27b sequence, especially its mature sequence, was conservative among different species. MiR-27b highly expressed in sheep skeletal muscle than other tissues. In skeletal muscle of Suffolk and Bashbay sheep, miR-27b was upregulated during foetal period and downregulated during postnatal period significantly (P<0.01), but it still kept a relatively higher expressionlevel in skeletal muscle of postnatal Suffolk sheep than Bashbay. There is a potential target site of miR-27b on 3'-UTR of sheep myostatin (MSTN) mRNA, and the double luciferase reporter assay proved that miR-27b could successfully bind on this site. When sheep satellite cells were in the proliferation status, miR-27b was upregulated and MSTN was downregulated significantly (P<0.01). When miR-27b mimics was transfected into sheep satellite cells, the cell proliferation was promoted and the protein level of MSTN was significantly downregulated (P<0.01). Moreover, miR-27b regulated its target gene MSTN by translation repression at an early step, and followed by inducing mRNA degradation in sheep satellite cells. Based on these results, we confirm that miR-27b could promote sheep skeletal muscle satellite cell proliferation by targeting MSTN and suppressing its expression.
pp 1119-1130 RESEARCH ARTICLE
The isolated and fragmented populations are highly susceptible to stochastic events, increasing the extinction risk because of the decline in putative adaptive potential and individual fitness. The population has high heterozygosity values and a moderate allelic diversity, the heterozygosity values are higher than in most other Crotalus species and snake studies. Possibly these high levels of genetic diversity can be related to a large founder size, high effective population size, multiple paternity and overlapping generations. We did not find the genetic structuring but the effective number of alleles (Ne) was 138.1. We found evidence of bottlenecks and the majority of rattlesnakes were unrelated, despite the small sample size, endemic status, the isolated and fragmented habitat. The genetic information provided in this study can be useful as a first approach to try to make informed conservation efforts for this species and also, important to preserve the habitat of this species; the endangered Abies–Pinus forest of the Nevado the Toluca Volcano.
pp 1131-1138 RESEARCH ARTICLE
Resistance to fatal disease bovine spongiformencephalopathy (BSE), due to misfolded prion protein in cattle, is associated with a 23-bp indel polymorphism in the putative promoter and a 12-bp indel in intron 1 of the PRNP gene. Gayal (Bos frontalis) is an important semiwild bovid species and of great conservation concern, but till today these indel polymorphisms have not been evaluated in gayals. Therefore, we collected 225 samples of gayals and evaluated the genetic indel polymorphism in the two regions of this PRNP gene. The results revealed high allelic frequencies of insertions at these indel sites: 0.909 and 0.667 for, respectively, the 23 bp and 12 bp indels, both also with significant genotype frequencies (χ2: 9.81; 23 bp and χ2: 43.56; 12 bp). At the same time, the haplotype data showed indel polymorphisms with extremely low deletion (0.01) in both regions of the PRNP gene. We compared these data with those reported for healthy and BSE-affected cattle (Bos taurus) breeds from two European countries, Germany and Switzerland, and significant difference (P <0.001) was observed between BSE-affected as well as the healthy cattle. Further, our datawere also extensively compared with previous reports on BSE and highly significant (P <0.001) outcomes were observed. This result suggested negligible genetic susceptibility to BSE in gayals. To the best of our knowledge, this study is the first comprehensive deciphering information about the PRNP indel polymorphisms of 23 bp and 12 bp in gayals, a semiwild species of China.
pp 1139-1146 RESEARCH ARTICLE
Crepidium acuminatum (Orchidaceae) is a threatened medicinal orchid that grows under shady and moist forest floor where light remains for a very short period of time. Mycorrhizal association is known to be essential for seed germination and seedling establishment in amajority of orchids. Identification of fungi that form mycorrhizae with orchids is of crucial importance for orchid conservation. We used both morphological as well as molecular approaches to study this plant–fungal interaction. Scanning electron microscopy showed that fungi grow and proliferate in the middle layers of the cortex. Also, spiral-root hairs were foundalong with root hairs, which is an unusual observation. Spiral-root hairs provide more surface area for fluid absorption and entrance of colonizers. Further, total root genomic DNA was isolated and fungal internal-transcribed spacer (ITS) regions were polymerase chain reaction (PCR)-amplified using specific primer combinations ITS1F/ITS4 and ITS1/ITS4tul. ITS sequences were obtainedand analysed to know the closest sequence matche in the GenBank using BLASTn hosted by NLM-NCBI. Subject sequences were identified to be belonging to three main genera, namely, Tulasnella, Aspergillus and Penicillium. Results indicate that mycorrhizal association is necessary for the growth and development of the plant. In addition, this symbiosis influences the distribution and rarity of this medicinally valuable taxon. Specific fungal partners may lead to an enhanced seed germination rate and increased efficiency of nutrient exchange between both the partners. Hence, knowledge of mycorrhizal fungi is essential for future in vitro germination and seedling establishment programmes, because they rely on fungi for germination. Identification of mycorrhizal fungi can be usedfor orchid propagation and conservation programmes.
pp 1147-1154 RESEARCH ARTICLE
Mycobacterium avium subsp. paratuberculosis (MAP) causes major problem in a wide range of animal species. In ruminant livestock including cattle, it causes a chronic disease called Johne’s disease, or paratuberculosis (pTB) which is currently considered as potential zoonosis, causing Crohn’s disease in humans. MAP infection susceptibility is suspected to be controlled by host genetics. Thus, selecting individuals according to their genetic structure could help to obtain bovine populations that are increasingly resistant to MAP infection. The aim of the present work was to investigate the association between toll-like receptor (TLR) 1 (+1380 G/A), TLR1 (+1446 C/A), TLR4 (+10 C/T), TLR9 (+1310 G/A) and solute carrier family 11 member 1 (SLC11A1) (+1066 C/G) mutations and MAP infection status in 813 cattle comprising East Anatolian Red crossbred, Anatolian Black crossbred and Holstein breed. TLR1 (+1380 G/A) mutation showed an association with bovineMAP (P<0.05). For the TLR1 (+1380 G/A) locus, the odds ratio for AG and AA genotypes versus GG genotypes were 2.31 (1.24–4.30; 95% confidence interval (CI)) and 0<0.001 (<0.001 to >999.999; 95% CI) which indicated that a proportion of AG homozygote was significantly higher in pTB-affected animals as compared with the control. General linear model analysis demonstrated higher MAP antibody response in TLR1 (+1380 AG) genotype as compared with TLR1 (+1380 GG) (P<0.0001). Present findings suggest that selection against TLR1 (+1380 G/A) may reduce the risk of pTBin bovine herds.
pp 1155-1168 RESEARCH ARTICLE
To analyse the mechanism of tumourigenic transformation of NIH3T3 cells at the transcriptional level, we used cancerogen 3-methylcholanthrene (3-MCA) and cancerogenic substance phorbol-12-myristate-13-acetate (PMA) to transform NIH3T3 cells and the assessment of transformation was performed using Giemsa staining and methylcellulose colony formation assay. Changes in gene expression profile were detected by MouseGenome 430 2.0 microarray; and quantitative real-time polymerase chain reaction and Western blotting were used to verify the expression changes of mRNAs and proteins, respectively. With the aidof bioinformatics method, five signalling pathways were identified to participate in different stages of NIH3T3 cell transformation. Further, our study suggested that oncogenes Cyclin A, Myc, Jun and the tumour suppressor gene Ppm1l may play important roles in these pathways.
pp 1169-1177 RESEARCH ARTICLE
Nonsyndromic hypodontia is a congenital absence of less than six permanent teeth, with a most common subtype maxillary lateral incisor agenesis (MLIA). Mutations in several genes have been described in severe tooth agenesis. The aim of this study was to search for the variants in wingless-type MMTV-integration site family member (WNT10A), paired box 9 (PAX9) and axis inhibitor 2 (AXIN2) genes, and investigate their potential role in the pathogenesis of non-syndromic hypodontia. Clinical examination and panoramic radiograph were performed in the cohort of 60 unrelated Slovak patients of Caucasian origin with nonsyndromic hypodontia including 37 MLIA cases and 48 healthy controls. Genomic DNA was isolated from buccal swabs and Sanger sequencing of WNT10A, PAX9 and AXIN2 was performed. Altogether, we identified 23 single-nucleotide variants, of which five were novel. We have found three rare nonsynonymous variants in WNT10A (p.Gly165Arg; p.Gly213Ser and p.Phe228Ile) in eight (13.33%) of 60 patients. Analysis showed potentially damaged WNT10A variant p.Phe228Ile predominantly occurred only in MLIA patients, and with a dominant form of tooth agenesis (odds ratio (ORdom) = 9.841; P = 0.045; 95% confidence interval (CI) 0.492–196.701;ORrec = 0.773; P = 1.000; 95% CI 0.015–39.877). In addition, the WNT10A variant p.Phe228Ile showed a trend associated with familial nonsyndromic hypodontia (P = 0.024; OR= 1.20; 95% CI 0.97–1.48). After Bonferroni correction, these effects remained with borderline tendencies. Using a 3D WNT10A protein model, we demonstrated that the variant Phe228Ile changes the proteinsecondary structure. In PAX9 and AXIN2, common variants were detected. Our findings suggest that the identified WNT10A variant p.Phe228Ile could represent risk for the inherited nonsyndromic hypodontia underlying MLIA. However, further study in different populations is required.
pp 1179-1183 RESEARCH ARTICLE
Pteronotus psilotis, a mormoopid bat, is an insectivorous, gregarious and strict cave-dwelling species that is found areas between the sea level and an elevation of about 1000 masl. This species is present in diverse habitats ranging from rain forest to dry deciduous forest. Nine microsatellite loci were developed for Wagner’s mustached bat, Pteronotus psilotis using the next-generation sequencing approach, and their utility for population genetics studies was assessed. All loci were polymorphic (7–15 alleles) and characterized in 30 individuals from three P. psilotis populations, with the levels of observed and expected heterozygosity ranging from 0.280 to 0.867 and 0.584 to 0.842, respectively. One locus showed significant departures from Hardy–Weinberg expectations after Bonferroni correction. Cross-amplification in 11 other bat species was tested, for which eight microsatellites were successfully amplified, and of these seven were polymorphic. The development of these new microsatellite loci will contribute to investigations of genetic population structure, genetic diversity and gene flow in P. psilotis populations, as well as in other closely related bat species.
pp 1185-1194 RESEARCH ARTICLE
Triple negative breast cancer (TNBC) is typically associated with poor and interindividual variability in treatment response. Cytochrome P450 family 1 subfamily B1 (CYP1B1) is a metabolizing enzyme, involved in the biotransformation of xenobiotics and anticancer drugs. We hypothesized that, single-nucleotide polymorphisms (SNPs), CYP1B1 142 C>G, 4326 C>G and 4360 A>G, and CYP1B1 mRNA expression might be potential biomarkers for prediction of treatment response in TNBC patients. CYP1B1 SNPs genotyping (76 TNBC patients) was performed using allele-specific polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism methods and mRNA expression of CYP1B1 (41 formalin-fixed paraffin embeddedblocks) was quantified using quantitative reverse transcription PCR. Homozygous variant genotype (GG) and variant allele (G) of CYP1B1 4326C>G polymorphism showed significantly higher risk for development of resistance to chemotherapy with adjusted odds ratio (OR): 6.802 and 3.010, respectively. Whereas, CYP1B1 142 CG heterozygous genotype showed significant association with goodtreatment response with adjusted OR: 0.199. CYP1B1 142C-4326G haplotype was associated with higher risk for chemoresistance with OR: 2.579. Expression analysis revealed that the relative expression of CYP1B1 was downregulated (0.592) in cancerous tissue compared with normal adjacent tissues. When analysed for association with chemotherapy response, CYP1B1 expression was found to be significantly upregulated (3.256) in cancerous tissues of patients who did not respond as opposed to those of patients who showed response to chemotherapy. Our findings suggest that SNPs together with mRNA expression of CYP1B1 may be useful biomarkers to predict chemotherapy response in TNBC patients.
pp 1195-1204 RESEARCH ARTICLE
A DNA region in the mitochondrial genome of the fungus Phycomyces blakesleeanus (Mucorales, Mucoromycota) was characterized in a population of wild-type strains. The region encodes a predicted protein similar to the reverse transcriptases encoded by mitochondrial retroplasmids of Neurospora species and other Sordariomycetes (Ascomycota), but is uncommon in other fungi. DNA sequences of this element, named mystique, are highly variable between the strains, having greater than 2.5% divergence, yet most of the nucleotide differences fall in codon positions that do not change the amino acid sequence. The high proportion of polymorphisms coupled to the rarity of nonsynonymous changes suggests that mystique is subject to counteracting forces of hypermutation and purifying selection. However, while evidence for negative selection may infer that the element provides a fitness benefit, some strains of P. blakesleeanus do not have the element and grow equivalently well as those strains with it. A mechanism to explain the variability between the mystique alleles is proposed, of error-prone replication through an RNA intermediate, reverse transcription and reintegration of the element into the mitochondrial genome.
pp 1205-1212 RESEARCH ARTICLE
Amphibian secretion is an important source of bioactive molecules that naturally protect the skin against noxious microorganisms. Collectively called antimicrobial peptides (AMPs), these molecules have a wide spectrum of action, targeting viruses, bacteria and fungi. Like many membrane and secreted proteins, AMPs have cleavable signal sequences that mediate and translocate the nascent polypeptide chains into the endoplasmic reticulum. Although it is accepted that the signal peptides (SPs) are simple and interchangeable, there is neither sequence nor structure that is conserved among all gene families. They derived from a common ancestor but developed different traits as they adapt to distinct environmental pressures. The aim of this study was to provide anoverview of the diversity of SPs of the frog, taking into account reported cDNA sequences and the evolutionary relationship among them. We analysed more than 2000 records that reported the relative abundance, diversity and evolutionary divergence based on the peptide signals of frog AMPs. We conclude that the physical properties of the sequence are more important than the specific peptidesin AMP SPs. Since there is significant overlapping among related genera, differences in secretion from different peptide types should be regulated by additional levels, such as posttranscriptional modifications or 5-UTR sequences.
pp 1213-1223 RESEARCH ARTICLE
Polycystic ovary syndrome (PCOS) is a common endocrine disorder in females, and is associated with altered metabolic processes in particular insulin resistance and diabetesmellitus. PCOS shares with type-2 diabetes (T2D) a number of features, including beta cell dysfunction, impaired glucose tolerance and dyslipidaemia. Recently, genomewide association studies (GWAS) have reported a number of genes with reproducible associations and susceptibilities to T2D. To address this, we examined the association between the T2D GWAS candidate genes (CDKAL1, CDKN2B, COL8A1, HHEX, IGF2BP2, KCNJ1, KCNQ1 and SLC30A8) and PCOS in Saudi women. A case–control study, includes 162 cases and 162 controls was enrolled. Genotyping was carried out by the allelicdiscrimination method. Our results showed that the variants including rs792837 of COL8A1, rs61873498 of KCNQ1 and rs13266634 of SLC30A8 genes to be significantly more frequent in PCOS patients than in controls. Our results suggest that COL8A1, KCNQ1 and SLC30A8, which are previously identified through GWAS as T2D-associated genes, are associated with PCOS.
pp 1225-1239 RESEARCH ARTICLE
Jasminum L. (Oleaceae) consists of ∼200 species that are distributed in tropical, subtropical and temperate regions of the world. In India, this genus is represented by ca 47 species of which 16 are endemic. Based on the nuclear (internal-transcribed spacer (ITS) region of nrDNA and chloroplast markers (matK, trnL-F and trnH-psbA), phylogenetic relationships in 22 species including one variety of Jasminum in India have been assessed. Maximum likelihood and Bayesian analyses from individual markers, as well as from combined dataset, reveal that the group is monophyletic if Menodora spp. are excluded from the analyses. Our analyses recovered three strongly supported clades. Ancestral character state reconstruction of taxonomically useful characters (leaf forms, leaf arrangement and flower colour) which were used to demarcate sections within the genus reveals homoplasy. Our study suggests that after split from the last common ancestor, there have been at least four reversals to unifoliolate condition. Pinnately compound leaf form evolved at least twice and trifoliolate condition evolved one time only. Alternate leaf form evolved at least twice, once inclade 1 and once in clade 3 and all the time from ancestors having opposite leaf forms. Flower colour evolution clearly depicts that clade 1 is yellow-flowered and clades 2 and 3 have admixture of white and yellow-flowered Jasminum species. Our study suggests that yellow-flowered condition evolved from the white-flowered ancestor. The present study is first to estimate the evolutionary history of Indian Jasmines.
pp 1241-1251 RESEARCH ARTICLE
Downy mildew (DM) caused by Sclerospora graminicola is the most calamitous disease of pearl millet. Therefore, for introgression of DM resistance (DMR) in HHB 197 (MH-1302), an elite pearl millet hybrid, a marker-assisted breeding was undertaken by targeting three DMR loci on linkage groups (LGs) 1, 2 and 4. Breeding programme was initiated by crossing HBL 11 (DM susceptible), male parent of HHB 197 hybrid with ICMP 451 (DM-resistant) to produce true F1 plants. By conducting three rounds of backcrossing and selection, BC3F1 lines were generated. Foreground selection was employed using six polymorphic simple sequence repeat (SSR) markers of the 18 total selected markers. Four of these markers were linked to LG 1, five to LG 2 and nine to LG 4. Background selection was performed in BC3F1 generation using 33 polymorphic SSR markers of a total of 56 evenly spread SSR markers in the pearl millet genome to check recovery of recurrent parent genome. On the basis of genotypic selection (foreground as well as background) using selected SSR markers, agronomic performance in field and DM screening in greenhouse; 10 improved HBL 11 lines were selected and crossed with ICMA 97111 to produce DM-resistant HHB 197 hybrid versions. Six putatively improved HHB 197 hybrids were successfully tested in first year trials at Hisar and Bawal locations of Haryana and two selected versions with higher yield and zero DM incidence will be further tested in multilocation trials.
pp 1253-1261 RESEARCH ARTICLE
Formulations from the traditional Indian medicine, Ayurveda, have long been considered to have potent life-style enhancing effects, possibly by their effect(s) on key life-history attributes. Although several studies have reported beneficial effects of these formulations on different components of life history, few have investigated their concurrent influence on various life-history traits. Here, we report the results of an investigation showing the effect of two well-known Ayurvedic formulations, Guduchi and Madhuyashti, on fecundity and longevity of Drosophila melanogaster. Flies were either grown (i.e., larval exposure) and/or maintained (i.e., adult exposure) on standard food supplemented with 0.5% Guduchi or 0.5% Madhuyashti. It was observed that the longevity of adult flies of both sexes was not affected on feeding Guduchifood, but fecundity of the females was greatly enhanced. Fecundity was also found to be affected by the adult food and whether their mates were grown on Guduchi or normal food. Madhuyashti, on the other hand, significantly reduced mean longevity and had a stimulatory effect on female fecundity. This fecundity enhancing effect however seemed to be mediated through its effect on the males. Interestingly, much of these effects interacted with age in a complex way, making it difficult to generalize the overall effect of these formulations on the reproductive output of the flies. Ourstudy underlines the importance of evaluating the interacting effects of these (and similar) formulations on a range of life-history traits in a holistic way to understand their utility better.
pp 1263-1279 RESEARCH ARTICLE
Identifying the best performing hybrid without a field test was essential to save resources and time. In this study, the genetic divergence was estimated using morphological and expressed sequence tag (EST)-derived simple sequence repeats (SSR) markers. Cluster analysis showed that APMS6A and RPHR 1005 belong to groups I and II, respectively, and the hybrid combination recorded the highest mean grain yield of 32.25 g among generated 40 F1s with standard heterosis of 8.4% over hybrid check, KRH2. The coefficient of marker polymorphism (CMP) value was calculated based on EST-SSRmarkers; it ranged from 0.40 to 0.80, and a higher CMP value of 0.80 was obtained for the parental combination APMS6A × RPHR1005. We predicted heterosis for 40 F1s based on correlation between CMP and standard heterosis in different traits with standard varietal and hybrid checks indicating positive correlation and significant value for grain yield per plant (r = 0.58**),productivity per day (r = 0.54**), productive tillers (r = 0.34*) and panicle weight (r = 0.42**). This study revealed that the relationship of molecular marker heterozygosity, along with the combining ability, high mean value of different traits,grouping of parental lines based on morphological and molecular characterization is helpful to identify heterotic patterns in rice.
pp 1281-1288 RESEARCH ARTICLE
APETALA1 (AP1), a floral meristem identity gene controls the flowering time and floral transition, and plays an important role in inflorescence and floral organ development. The full-length cDNA for AP1 was obtained by rapid amplification ofthe cDNA ends (RACE) so that the roles of AP1 in Lonicera macranthoides (Lm-AP1) could be better understood. AP1 (accession number in GenBank: MF418642) consisted of a 729-bp open reading frame encoding a protein that contained 242 amino acids, had a deduced molecular mass of 27.9919 kDa and a theoretical isoelectric point of 8.75. No signal peptide or transmembranedomains were detected in the sequences located in the nucleus, but it contained conserved sequences for MADS and the K-box. In the secondary structure, the alpha helix accounts for 60.74%, the beta turn 3.72%. The real-time polymerase chain reaction revealed that AP1 was more highly expressed in flowers, especially at the fourth flowering stage, which implied that it may play a role in flower development. Other L. macranthoides organs, such as stems and leaves, also expressed AP1. This research provided the basis for further analysis of the AP1 functional mechanism during L. macranthoides development.
pp 1289-1294 RESEARCH ARTICLE
Gestational diabetes mellitus (GDM) represents a common carbohydrate metabolism disorder during pregnancy. The objective of this study was to evaluate the expression levels of mitofusin 2 (MFN2) expression in placentae of GDM patients compared to that in the placental tissues from normal uncomplicated pregnancies. A total of 70 subjects were enrolled from September 2014 to June 2016, including 42 patients with GDM (the GDM group) and 28 normal uncomplicated pregnancies (the control group). Immunohistochemical staining and qRT-PCR were used for the detection of the expression levels and distribution of MFN2 in the placentae of GDM patients and normal controls. Kolmogorov–Smirnov test was used for statistical analysis. P < 0.05 and P < 0.01 were used for assessing statistical significance. The baseline characteristics were comparable in both groups. The 1-h and 2-h postprandial glucose levels (PPG) were 7.94±1.26 versus 6.88±0.51 mmol/L and 7.01±1.34 versus 6.14±0.63 mmol/L, respectively,for the GDM group and the control group (P < 0.05). The relative expression levels of MFN2 mRNA were 0.982±1.242 for GDM and 1.257 ± 0.815 for control, respectively, with significant between group difference (P < 0.01). Immunohistochemical staining analysis showed that MFN2 was mostly distributed in the cytoplasm of syncytiotrophoblasts under optical microscopy. Additionally,about 50% of samples of the GDM group were within the intensity of moderate staining of MFN2 and more than 50% of patients in the control group were within the intensity of strong staining of MFN2. The expression levels of MFN2 in GDM placentae was significantly lower compared to that of placentae from normal uncomplicated pregnancies.
pp 1295-1306 RESEARCH ARTICLE
Genomic simple sequence repeat (SSR) markers were used to fingerprint and determine genetic similarity (GS) of the watermelon breeding lines, as well as the purity of their hybrid derivatives. Cluster analysis and Jaccard’s distance coefficients using the unweighted pair group method with arithmetic mean (UPGMA) have classified these lines into three major groups. Notwithstanding, the genetic background of these lines is narrow as revealed by the restricted GS coefficients. Fifty-five sets of SSR markers were employed in this study. Fourteen of these markers were polymorphic between the breeding lines and were used for assessing hybrid purity. Cross-checking assay validated nine SSR markers as informative SSR markers for purity detection of these hybrids. To confirm the accuracy and efficiency of these markers, their derived PCR products were further sequenced, and ClSSR09643, ClSSR18153 and ClSSR01623 were selected as high-efficiency SSR markers. Interestingly, SSR markers ClSSR09643 and ClSSR18153 were broadly applied for purity detection of more than two different hybrids, while SSR marker ClSSR01623 behaved as a specific marker forpurity detection in this study. Genetic purity of six commercial watermelon hybrids was definitely evaluated using these SSR markers. Genetic purity of all tested hybrids exceeded 96% while the field purity was above 98%. Genetic purity test was an emergency for identifying off-types and selfed female in a lot of hybrid seeds. Here, we elucidated the potential of nine SSR markers including threewith higher breeding selection efficiency. We recommended them to seed company for purity improvement of watermelon commercial hybrid varieties.
pp 1307-1313 RESEARCH ARTICLE
Some of the single-nucleotide polymorphisms in miRNA genes have been studied to date to find their association with the risk of breast cancer (BC). However, no study has been conducted to investigate the association of the mir-520f rs75598818G>A in BC. In the present study, rs75598818 association with BC in an Iranian population has been investigated, and an in silico analysis was performed to predict the function of rs75598818 polymorphism in BC. The rs75598818 was genotyped in 129 BC patients and 144 healthy women, using the PCR-RFLP method. The frequency of alleles and genotypes were considered to find the associations between rs75598818 alleles/genotypes, and BC risk and pathological characteristics of the patients. Statistical analysis showed that the rs75598818 GA genotype was significantly associated with BC (GA versus GG, OR=0.50, 95% CI: 0.25–0.98, P =0.041), highstage BC (stage III/IV versus I/II, GA versus GG, OR=0.27, 95% CI: 0.09–0.81, P =0.015), and HER-2 positive status (GA versus GG, OR=19.00, 95% CI: 4.64–77.82, P <0.001). Notably, the rs75598818 GA genotype has a negative association pattern since it reduces the risk of BC and high stage BC. Conversely, it increases the risk of HER-2 positivity. Computational results suggested that the rs75598818 polymorphism affects the stability of mir-520f stem-loop and as a result miR-520f-3p production that is a potential tumour suppressor. A contribution of the mir-520f rs75598818 polymorphism to BC had been unexplored before. In the present study, we performed an association study and a bioinformatics approach to evaluate this polymorphism in BC. However, further functional experiments and large-scale association studies with various ethnicities are required to elaborate our findings.
pp 1315-1325 RESEARCH ARTICLE
ASHLEY N. TURNER REAGAN S. ANDERSEN IVY E. BOOKOUT LAUREN N. BRASHEAR JAMES C. DAVIS DAVID M. GAHAN JOHN P. GOTHAM BARAA A. HIJAZ ASHISH S. KAUSHIK JORDAN B. MCGILL VICTORIA L. MILLER ZACHARIAH P. MOSELEY CERISSA L. NOWELL RIDDHI K. PATEL MIA C. RODGERS YAZEN A. SHIHAB AUSTIN P. WALKER SARAH R. GLOVER SAMANTHA D. FOSTER ANIL K. CHALLA
Nodal-related protein (ndr2) is amember of the transforming growth factor type β superfamily of factors and is required for ventral midline patterning of the embryonic central nervous system in zebrafish. In humans, mutations in the gene encoding nodal cause holoprosencephaly and heterotaxy. Mutations in the ndr2 gene in the zebrafish (Danio rerio) lead to similar phenotypes, including loss of the medial floor plate, severe deficits in ventral forebrain development and cyclopia. Alleles of the ndr2 gene have been useful in studying patterning of ventral structures of the central nervous system. Fifteen different ndr2 alleles have been reported in zebrafish, of which eight were generated using chemical mutagenesis, four were radiation-induced and the remaining alleles were obtained via random insertion, gene targeting (TALEN) or unknown methods. Therefore, most mutation sites were random and could not be predicted a priori. Using the CRISPR-Cas9 system from Streptococcus pyogenes, we targeted distinct regions in all three exons of zebrafish ndr2 and observed cyclopia in the injected (G0) embryos.We show that the use of sgRNA-Cas9 ribonucleoprotein (RNP) complexes can cause penetrant cyclopic phenotypes in injected (G0) embryos. Targeted polymerase chain reaction amplicon analysis using Sanger sequencing showed that most of the alleles had small indels resulting in frameshifts. The sequence information correlates with the loss of ndr2 activity. In this study, we validate multiple CRISPR targets using an in vitro nuclease assay and in vivo analysis using embryos. We describe one specific mutant allele resulting in the loss of conserved terminal cysteine-coding sequences. This study is another demonstration of the utility of the CRISPR-Cas9 system in generating domain-specific mutations and provides further insights into the structure–function of the ndr2 gene.
pp 1327-1337 RESEARCH ARTICLE
The phenotypic and microsatellite marker information of nine strains of catla (Catla catla) for growth trait was used to infer relationship within and between strains. This information helped in optimizing the proportion of individuals to be used from each strain while creating a base population for selective breeding. For this purpose, nine strains were collected from different sources and places of India namely West Bengal, Bihar, Odisha, Andhra Pradesh and Uttar Pradesh. Two riverine sources i.e. Ganga and Subarnarekha were also represented among the nine strains collected for base population. They were brought to Indian Council of Agricultural Research-Central Institute of Freshwater Aquaculture (ICAR-CIFA) at fry stage and reared separately till fingerlings. After passive integrated transponder tagging fingerlings were stocked in three communal ponds for one year culture. Live body weights were then measured and least square means were obtained after pond effect correction. A wide range of variation was observed among and between strains. Microsatellite markers were used to estimate genetic differences of different strains of catla using pair wise FST estimates. Overall multi locus FST, including all loci was estimated to be 0.4137 (P < 0.05), indicating genetic heterogeneity among them. Analysis of molecular variance revealed that, 58.63% of variation was due to within individual variation, 3.45% of variation was due to among individuals within strain and 37.92% was due to among strain variations. Both phenotypic as well as microsatellite data will be used to form a base population of catla with individuals from the stock having broad genetic variation for selective breeding programme.
pp 1339-1351 RESEARCH ARTICLE
The American cranberry (Vaccinium macrocarpon) is an endemic domesticated species that has become an economically important commercial fruit crop. The USDA-ARS National Clonal Germplasm Repository (NCGR) houses the national Vaccinium collection, which includes representatives of historical cranberry cultivars and wild-selected germplasm. The objective of this study wasto examine the genotypes of 271 cranberry plants from 77 accessions representing 66 named cultivars using 12 simple-sequence repeats to assess clonal purity and cultivar relatedness. Using principal components analysis and neighbour-joining based on estimated genetic distances between individuals, we identified 64 unique genotypes and observed that intracultivar variants (i.e. subclones) existed in the germplasm collection and in the commercial bogs where some accessions originated. Finally, through a comparison of the genotypes of this study with the previous studies, pedigree analysis and the study of the geographic distribution of cranberry diversity, we identified consensus genotypes for many accessions and cultivars. We highlight the important role that the NCGR collection playsfor ex situ conservation of cranberry germplasm for future breeders and researchers. The NCGR continues to search for historically relevant cultivars absent from the collection in an effort to preserve these genotypes before they are lost and no longer commercially grown.
pp 1353-1361 RESEARCH ARTICLE
The receptor-like kinase ERECTA (ER) plays vital roles in plant adaptation under environmental stress including shade avoidance in Arabidopsis. In our previous study, we identified four ER paralogues in soybean (GmERs) that showed high similarities to Arabidopsis ER. Each GmER was predicted to generate diverse alternative splicing variants. However, whether soybean GmERs contribute to shade avoidance is unknown. Here we report our characterization of GmERs in response to shading. Promoter::GUS staining analysis shows that expression of GmER paralogous genes was differentially induced under shade stress. Further analyses show that GmERa.1 and GmERa.2 exhibit a larger distinction in length than the other GmER variants. GmERa.2 has the shortest length of amino acid with only 15 leucine-rich repeats which is the part of the extracellular domain of GmERa.1. Overexpression of GmERa.2 fully rescued the hypocotyl length, leaf area and petiole length, and the sensitivity of the hypocotyl of Arabidopsis mutant er-3 to shading, suggesting that the truncated extracellular domain of GmERa might contribute importantly to shade avoidance.
pp 1371-1378 RESEARCH ARTICLE
There is a drastic increase in the number of people suffering from coronary artery disease (CAD) worldwide with Indians being no exception. Being a developing country and experiencing a dramatic shift in lifestyle and eating habits, urbanization and industrialization, all these factors have collectively predisposed the Indian population towards CAD and the prevalence data arequite alarming. Genetic studies have disclosed the role of genes in CAD susceptibility and severity. One such gene is proprotein convertase subtilisin/kexin type 9 (PCSK9) which is sought to modulate the cholesterol levels and hence, has implications in CAD. We aim to explore the association of PCSK9 A/G (rs505151) polymorphism and hence, the susceptibility towards CAD in the north Indian population. Five-hundred angiographically confirmed CAD patients and 500 healthy individuals as control were genotyped by polymerase chain reaction-restriction fragment length polymorphism. Statistical analysis revealed a significant association with the G allele with odds ratio (OR)=1.50, 95% confidence interval (CI)=1.22–1.85 and P=0.000. Also, a strong association was observedfor CAD risk with OR=1.590, 95% CI=1.106–2.284 and P=0.012. However, the homozygous GG mutant genotype was found to be completely absent from our population. Analysis of the dominant model also revealed an association with CAD risk. Our work demonstrated for the first time the association of PCSK9 A/G (rs505151) polymorphism with CAD risk in the north Indian population.
pp 1379-1388 RESEARCH ARTICLE
Conyza blinii H. Lév., the most effective component is saponin, is a biennial medicinal material that needs to be overwintered. WRKY transcription factors family is a large protein superfamily that plays a predominant role in plant secondary metabolism, but their characteristics and functions have not been identified in C. blinii. The CbWRKY24 sequence was selectedfrom the transcriptome database of the C. blinii leaves constructed in our laboratory. Phylogenetic tree analysis revealed that it was associated with AaWRKY1 which can regulate artemisinin synthesis in Artemisia annua. Expression analysis in C. blinii revealed that CbWRKY24 was mainly induced by methyl jasmonate (MeJA) and cold treatments. Transcriptional activity assay showed that it had an independent biological activity. Overexpression of CbWRKY24 in transient transformed C. blinii resulted in improved totalsaponins content, which was attributed to upregulate the expression level of keys genes from mevalonate (MVA) pathway in transient transformed plants compared to wild type (WT) plants. Meanwhile, overexpression the CbWRKY24 in transient transformed tomato fruits showed that the transcript level of related genes in lycopene pathway decreased significantly when compared to WT tomatofruits. Additionally, the MeJA-response-element was found in the promoter regions of CbWRKY24 and the histochemical staining experiments showed that promoter had GUS activity in transiently transformed tobacco leaves. In summary, our results indicated that we may have found a transcription factor that can regulate the biosynthesis of terpenoids in C. blinii.
pp 1389-1406 RESEARCH ARTICLE
Identification of superior alleles for agronomic traits in genetic resources of oilseed rape (Brassica napus L.) would be useful for improving the performance of locally adapted cultivars in Iran. The objective of the present work was to analysethe genetic variation and inheritance of important agronomic traits in a doubled haploid population derived from a cross between two German oilseed rape cultivars, Sansibar and Oase. Field experiments were performed in 2016–2017 with 200 doubled haploid lines and the parental genotypes applying an alpha-lattice design with two replicates. Phenological traits were recorded duringthe cultivation period and at maturity, seed yield, yield components and seed quality traits were determined. Significant genetic variation was found in most of the traits and heritabilities ranged from medium (48.5%) for days to end of flowering to high (92.6%) for oil content. A molecular marker linkage map was used to map 36 QTL for different traits on 17 linkage groups. Betweenthree and four QTL were identified for each seed yield, seed weight, oil and protein content. Some of the plant material and positive QTL alleles identified for agronomic traits may be useful for improving those characters in locally adapted cultivars in Iran.
pp 1407-1412 RESEARCH ARTICLE
Type-2 diabetes (T2DM) is ametabolic disorder characterized by long-terminsulin resistance, impaired insulin secretion from β-cells, and loss of beta cell mass and function. Inflammation and oxidative stress play a key role in the development of diabetes and are associated with insulin resistance. Notably, recent studies have demonstrated an association between body iron stores, insulin resistance and T2DM. Free iron, a powerful pro-oxidant molecule, is involved in oxidative stress, lipid peroxidation and endothelial dysfunction via its ability to generate free radicals. Specifically, the accumulation of iron in beta cells triggers oxidative stress and DNA damage, which have been reported to be associated with β-cell death and apoptosis. Solute carrier family-11 member-2 (SLC11A2) functions to transport ferrous iron and some divalent metal ions throughout the plasmamembrane and across endosomalmembranes. Functional polymorphisms in the SLC11A2 gene have been reported to cause excess storage of iron, resulting in iron overload. In this study, we evaluated the association between T2DM and SLC11A2 gene variants IVS4+44C/A, 1303C/A and 1254T/C by performing PCR-RFLP analysis on 100 T2DM patients and 100 healthy subjects. PCR products were digested with MnlI, MboI and SfanI restriction endonucleases and the products were then separated by 3% agarose gel electrophoresis. The genotype frequencies of the 1254T/C and 1303C/A SLC11A2 gene variants did not differ between healthy controls and T2DM patients (P > 0.05). But, inrecessive model (P = 0.037) and homozygous CC genotype (P = 0.030) for IVS4+44C/A showed significant correlation with T2DM risk. It is thought that presence of C allele of IVS4+44C/A plays pathological roles.
pp 1413-1420 RESEARCH ARTICLE
When compared to C57BL/6J (B6) mice, DDD/Sgn (DDD) mice has substantially higher plasma insulin levels in both sexes. In this study, we performed quantitative trait loci (QTL) mapping of plasma insulin levels in F2 male mice produced by crosses between DDD and B6 mice. By single-QTL scans, we identified one significant QTL on chromosome 9. When body weight was included as an additive covariate, we identified two significant QTL on chromosomes 9 and 12; the latter coincided with a QTL that was previously identified in F2 female mice produced by the same two strains. The inheritance mode and the direction of the allelic effect of QTL on chromosome 12 were similar in both sexes, but those on chromosome 9 differed between males and females, suggesting that the QTL on chromosome 9 was sex-specific. Based on phenotypic correlations of plasma insulin levels with body weight and plasma levels of total cholesterol, triglyceride and testosterone, we subsequently assessed whether these insulin QTL explain the variation in other metabolic traits by using a point-wise significance threshold of P = 0.05. QTL on chromosome 12 had no significant effect on any trait. In contrast, QTL on chromosome 9 had significant effects on body weight and total cholesterol level. We postulate that Gpr68 and Cyp19a1 are plausible candidate genes for QTL on chromosomes 12 and 9, respectively. These findings provide insight into the genetic mechanisms underlying insulin metabolism.
pp 1421-1431 RESEARCH ARTICLE
Nonrandom segregation ratios of alleles ‘segregation distortion’ can have a striking impact on transmission genetics, and with widespread availability of genetic markers has been shown to be a frequent phenomenon. To investigate the possible effect of genetic interaction on segregation distortion and genetic map construction, the segregation and mapping of genetic markers locatedon wheat chromosomes 1A and 1B were followed in four recombinant substitution line (RSL) populations, produced using four chromosome-arm substitution lines (CASLs 1AS, 1AL, 1BS and 1BL) of wild emmer (Triticum turgidum var. dicoccoides, accession TTD140) in the background of the common wheat (T. aestivum) cultivar Bethlehem (BLH), each crossed to BLH itself. Using these four RSL populations, four genetic maps of chromosome 1 arms were constructed. A total of 22 genetic markers representing 19 loci were assigned to chromosome 1A, and 32 markers representing 30 loci were assigned to 1B. For chromosome 1B, two linkage maps were also constructed using RFLP data of an F2 population derived from the same cross combination as the RSLs. The RSL and F2 maps varied in genetic distances, but showed the same linear order of DNA markers. Segregation analysis revealed strong selection against BLH alleles on chromosome 1B, skewing the allelic frequency distribution in favour of TTD in both F2 and RSL populations at all marker loci. On the contrary, strong selection against TTD alleles on chromosome 1A was detected for some loci in the BLH × CASL1AL RSLs, and their distribution was significantly skewed to BLH. F2 populations always showed more segregation distortion than the corresponding RSLs. More markers near the region of chromosome 1B shared by both CASL1BS and 1BL (∼55 cM on chromosome 1B across the centromere) showed significantly distorted segregation in the BLH × CASL1BL population than in thecorresponding BLH × CASL1BS populations. Six markers located on chromosome 1A region shared by CASL1AS and 1AL showed significantly distorted segregation in 1AL-RSL, while no marker showed distorted segregation in 1AS-RSL. These results indicated that genetic factor(s) in the centromere region cause the distorted segregation of genetic markers on wheat chromosome 1B.
pp 1433-1444 RESEARCH ARTICLE
Eight candidate housekeeping genes were examined as internal controls for normalizing expression analysis of durum wheat (Triticum durum L.) under drought and salinity stress conditions. Quantitative real-time PCR was used to analyse gene expression of multiple stress levels, plant ages (24 and 50 days old), and plant tissues (leaf and root). The algorithms BestKeeper, NormFinder, GeNorm, the delta Ct method and the RefFinder were applied to determine the stability of candidate genes. Under drought stress, the most stable reference genes were glyceraldehyde-3 phosphate, ubiquitin and βtubulin2-, whereas under salinity stress conditions, eukaryotic elongation factor 1-α, glyceraldehyde-3 phosphate and actin were identified as the most stable reference genes. Validation with stress-responsive genes NAC29 and NAC6 demonstrated that the expression level of target genes could be determined reliably with combinations of up to three of the reference genes. This is the first report on reference genes appropriate for quantification of target gene expression in T. durum under drought and salt stresses. Results of this investigation may be applicable to other Triticum species.
pp 1445-1449 RESEARCH NOTE
Monosialotetrahexosylganglioside (GM1) is a rare lysosomal storage disorder caused by the deficiency of beta-galactosidase (β-Gal) encoded by galactose beta 1 (GLB1). It is clinically characterized by developmental delay attributed to multifold accumulation of GM1 gangliosides in nerve cells. In this study, we present a case of infantile GM1 gangliosidosis in a consanguineous Pakistani family. The child was presented with developmental delay, hepatosplenomegaly and recurrent chest infections at 7.5 months of age. Radiological and biochemical investigations including magnetic resonance imaging (MRI), bonemarrow biopsy and urine oligosaccharide analyses suggested lysosomal storage disorder. Significantly low levels of β-Gal enzyme confirmed the diagnosis of GM1 gangliosidosis. DNA sequencing of GLB1 identified a homozygous 2-bp deletion c.881-882delAT (p.Tyr294Terfs) in exon 8. In silico analysis supported the deleterious effect of the variant. This study extends GLB1mutation spectrum and should benefit genetic counselling and prenatal diagnosis of the affected family.
pp 1451-1456 RESEARCH NOTE
Genetics of resistance to Cercospora leaf spot disease caused by Cercospora canescens and Psuedocercospora cruenta in yardlong bean (Vigna unguiculata ssp. sesquipedalis) × grain cowpea (V. unguiculata ssp. unguiculata) populations
Yardlong bean (Vigna unguiculata ssp. sesquipedalis), a type of cowpea, is an important vegetable legume of Asia. Cercospora leaf spot (CLS) caused by Cercospora canescens and Psuedocercospora cruenta is an important phytopathological problem of the yardlong bean grown in tropical regions. The objectives of this study were to (i) determine mode of inheritance of resistance to CLS caused by C. canescens and P. cruenta, (ii) estimate the heritability of the resistance, (iii) estimate genetic effects on the resistance using six basic populations generated from the cross between the susceptible yardlong bean ‘CSR12906’ and the resistant grain cowpea (V.unguiculata spp. unguiculata) ‘IT90K-59-120’. Segregation for the resistance to both fungi in the F2 population fitted both 3 : 1 ratio and 13 : 3 ratio of susceptible:resistant, while that in the BC2 ((CSR12906×IT90K-59-120)×IT90K- 59-120) population fitted a 1 : 1 ratio, suggesting one recessive gene or two genes with inhibitory gene action control the resistance. Generation mean analysis showed that a simple additive–dominance model was adequate to explain the genetic control of CLS disease resistance, indicating that a single gene controls the resistance. The average number of major genes (effective factors) controlling the resistance was estimated to be 1.05 and 0.92 for C. canescens and P. cruenta, respectively. The broad-sense heritability calculated for resistance to both diseases was higher than 0.90. Altogether, these results indicated that the resistance to CLS disease caused by C. canescens and P. cruenta in grain cowpea IT90K-59-120 is a highly heritable trait governed by a single major recessive gene.
pp 1457-1461 RESEARCH NOTE
Asiatic wild dog (Cuon alpinus) or dhole is an endangered canid with fragmented distribution in South, East and Southeast Asia. The remaining populations of this species face severe conservation challenges from anthropogenic interventions, but only limited information is available at population and demography levels. Here, we describe the novel molecular approaches for unambiguous species and sex identification from noninvasively collected dhole samples. We successfully tested these assays on 130 field-collected dhole faecal samples from the Vidarbha part of central Indian tiger landscape that resulted in 97 and 77% successrates in species and sex identification, respectively. These accurate, fast and cheap molecular approaches prove the efficacy of such methods in gathering ecological data from this elusive, endangered canid and show their application in generating population level information from noninvasive samples.
pp 1463-1468 RESEARCH NOTE
Low-molecular-weight glutenin subunits (LMW-GSs) are one of the important factors for the dough processing quality. In this study, a novel LMW-GS, designated LMW-N13, from the wheat relative species Aegilops uniaristata PI 554421 was cloned and characterized. Unlike previously published LMW-GSs, LMW-N13 has a large molecular weight and is the largest LMW-GS published thus far. Sequence alignments demonstrated that LMW-N13 is a LMW-i-type subunit but contains nine cysteine residues which is one more than typical LMW-i-type subunits. In addition, four insertions are present in the repetitive domain that resulted in the large molecular weight. In vitro analysis showed that LMW-N13 could improve the dough quality of different base flours.
pp 1469-1472 RESEARCH NOTE
Hereditary spastic paraplegias are clinically and genetically heterogeneous degenerative disorders, and pathological variants in the autosomal recessive ZFYVE26 gene are considered as very rare causes. We describe a novel mutation in ZFYVE26 gene found in a patient with autosomal recessive spastic paraplegias. The use of a ‘target-gene’ approach allowed us to expand the clinical spectrum associated with hereditary spastic paraplegias.
pp 1473-1478 RESEARCH NOTE
We performed phylogenetic analyses of HBG genes to assess its origin and interspecific variation among primates. Our analyses showed variation in HBG genes copy number ranging from one to three, some of them pseudogenes. For platyrrhines HBG genes, phylogenetic reconstructions of flanking regions recovered orthologous clades with distinct topologies for 5' and 3' flanking regions. The 5' region originated in the common ancestor of platyrrhines but the 3' region had an anthropoid origin. We hypothesize that the platyrrhine HBG genes of 5' and 3' heterophyletic origins arose from subsequent fusions of the (earlier) platyrrhine 5' portion and the (later) anthropoid 3' portion.
pp 1479-1484 RESEARCH NOTE
Ocular albinism type 1 (OA1) is an X-linked inherited disease characterized by impaired visual acuity, congenital nystagmus, foveal hypoplasia, hypopigmentation of iris and fundus. It is caused by mutations in the G protein-coupled receptor143 (GPR143) gene. The genetic characteristics of OA1 have not been well defined in Asians. In this study, six members from three consecutive generations of a Korean family with OA1 were enrolled. We performed whole-exome sequencing followed by validation and segregation analysis. Two affected patients underwent detailed ophthalmic examinations and eye movement recordings. Of the two affected males, the proband had all classical phenotypes of OA1, but the other showed isolated foveal hypoplasia without nystagmus. We identified a hemizygous missense (c.623C > A, p.Ala208Glu) mutation of GPR143 in affected males. This mutation was also present as heterozygous in two obligate female carriers, and was not found in unaffected members. Our data expands thespectrum of phenotypes and genotype in GPR143 in Asians, and highlights the phenotypic heterogeneity in OA1.
pp 1485-1491 REVIEW ARTICLE
Genetic variation of language genes affect neurophysiology of brain and can thus influence the way people respond to environmental language input, leading to differences in terms of their response to environmental language learning. Conversely, language learning environment too can affect gene expressions through neuroepigenetic mechanisms, leading to increasedinterindividual differences. Further, language-related cognitive processes such as learning, working memory and perception; and language-related affective factors such as stress and positive emotion involve neuroplasticity, which is also epigenetically regulated. Language intervention methods must understand the extent and the type of difficulties, and must offer personalized learning andmedical solutions. Medical intervention in terms of epigenetics and neurotransmitter regulation is proposed in addition to effectiveteaching methods to aid in effective language acquisition
pp 1493-1507 REVIEW ARTICLE
Ulcerative colitis (UC), one of the two clinical subtypes of inflammatory bowel disease is perceived as a potential 'sleeping giant' in the Indian subcontinent. Clinical manifestation is overall believed to be the same across ethnic groups but overwhelming genetics from large European and fewer non-European studies have revealed shared as well as unique disease susceptibly signaturesbetween them, pointing to population specific differences at genomic and environmental levels. A systematic recount of the four major eras in UC genetics spanning earliest linkage analysis, cherry picked candidate gene association studies, unbiased genomewide association studies, their logical extension in trans-ethnic setting (Immunochip study), lastly whole exome sequencing efforts forrare variant burden; and lessons learnt thereof in context of genetically distinct Indian population was attempted in this review. Genetic heterogeneity manifesting at allelic/locus level across these approaches has been the consistent finding through the range of pan India studies. On the other hand, these salient findings also highlight the limitations of even the best of these genetic leadsfor prognostic/clinical application. The imminent need, therefore, for the UC research community to adopt newer approaches/tools with improved study design to (i) gain better insight into genetic/mechanistic basis of disease; (ii) identify biomarkers of immediate translational value; and (iii) develop new/alternate therapeutic options is emphasized at the end.