Salinity severely limits wheat production in many parts of the world. Development of salt tolerant varieties represents the most practical option for enhancing wheat production from these areas. Application of marker assisted selection may assist in fast tracking development of salt tolerant wheat varieties. However, SSR markers available in the public domain are not specifically targeted to functional regions of wheat genome, therefore large numbers of these need to be analysed for identification of markers associated with traits of interest. With the availability of a fully annotated wheat genome assembly, it is possible to develop SSR markers specifically targeted to genic regions. We performed extensive analysis to identify candidate gene based SSRs and assessed their utility in characterizing molecular diversity in a panel of wheat genotypes. Our analysis revealed, 161 SSR motifs in 94 salt tolerance candidate genes of wheat. These SSR motifs were nearly equally distributed on the three wheat sub-genomes; 29.8% in A, 35.7% in B and 34.4% in D sub-genome. The maximum number of SSR motifs was present in exons (31.1%) followed by promoters (29.8%), 5’UTRs (21.1%), introns (14.3%) and 3’UTRs (3.7%). Out of the 65 candidate gene based SSR markers selected for validation, 30 were found polymorphic based on initial screening and employed for characterizing genetic diversity in a panel of wheat genotypes including salt tolerant and susceptible lines. These markers generated an average of 2.83 alleles/locus. Phylogenetic analysis revealed four clusters. Salt susceptible genotypes were mainly represented in clusters I and III, whereas high and moderate salt tolerant genotypes were distributed in the remaining two clusters. Population structure analysis revealed two sub-populations, sub-population 1 contained the majority of salt tolerant whereas sub-population 2 contained majority of susceptible genotypes. Moreover, we observed reasonably higher transferability of SSR markers to related wheat species. We have developed salt responsive gene based SSRs in wheat for the first time. These were highly useful in unravelling functional diversity among wheat genotypes with varying responses to salt stress. The identified gene based SSR markers will be valuable genomic resources for genetic/association mapping of salinity tolerance in wheat.
Wheat leaf rust caused by Puccinia triticina Eriks is an important disease that causes yield losses of up to 40% in susceptible varieties. Tetraploid emmer wheat (T. turgidum ssp. Dicoccum), commonly called Khapli wheat in India, is known to have evolved from wild emmer (Triticum turgidum var. dicoccoides), and harbors a good number of leaf rust resistance genes. In the present study, we are reporting on the screening of one hundred and twenty-three dicoccum wheat germplasm accessions against the leaf rust pathotype 77-5. Among these, an average of 45.50% of the germplasms were resistant, 46.74% were susceptible, and 8.53% had mesothetic reactions. Further, selected germplasm lines with accession numbers IC138898, IC47022, IC535116, IC535133, IC535139, IC551396, and IC534144 showed high level of resistance against the eighteen prevalent pathotypes. The infection type varied from “;”, “;N”, “;N1” to “;NC”. PCR-based analysis of the resistant dicoccum lines with SSR marker gwm508 linked to the Lr53 gene, a leaf rust resistance gene effective against all the prevalent pathotypes of leaf rust in India and identified from a T. turgidum var. dicoccoides germplasm, indicated that Lr53 is not present in the selected accessions. Moreover, we have also generated 35K SNP genotyping data of seven lines and the susceptible control, Mandsaur Local, to study their relationships. The GDIRT tool based on homozygous genotypic differences revealed that the seven genotypes are unique to each other and may carry different resistance genes for leaf rust.
Cadmium (Cd) is a hazardous heavy metal, toxic to our ecosystem even at low concentrations. Cd stress negatively affects plant growth and development by triggering oxidative stress. Limited information is available on the role of iron (Fe) in ameliorating Cd stress tolerance in legumes. This study assessed the effect of Cd stress in two lentil (Lens culinaris Medik.) varieties differing in seed Fe concentration (L4717 (Fe-biofortified) and JL3) under controlled conditions. Six biochemical traits, five growth parameters, and Cd uptake were recorded at the seedling stage (21 days after sowing) in the studied genotypes grown under controlled conditions at two levels (100 μM and 200 μM) of cadmium chloride (CdCl2). The studied traits revealed significant genotype, treatment, and genotype × treatment interactions. Cd-induced oxidative damage led to the accumulation of hydrogen peroxide (H2O2) and malondialdehyde in both genotypes. JL3 accumulated 77.1% more H2O2 and 75% more lipid peroxidation products than L4717 at the high Cd level. Antioxidant enzyme activities increased in response to Cd stress, with significant genotype, treatment, and genotype × treatment interactions (p < 0.01). L4717 had remarkably higher catalase (40.5%), peroxidase (43.9%), superoxide dismutase (31.7%), and glutathione reductase (47.3%) activities than JL3 under high Cd conditions. In addition, L4717 sustained better growth in terms of fresh weight and dry weight than JL3 under stress. JL3 exhibited high Cd uptake (14.87 mg g−1 fresh weight) compared to L4717 (7.32 mg g−1 fresh weight). The study concluded that the Fe-biofortified lentil genotype L4717 exhibited Cd tolerance by inciting an efficient antioxidative response to Cd toxicity. Further studies are required to elucidate the possibility of seed Fe content as a surrogacy trait for Cd tolerance.
Teosinte is the closest wild ancestor of maize and is used as a valuable resource for taxonomical, evolutionary and genetic architectural studies of maize. Teosinte is also a repository of numerous diverse alleles for complex traits, including nutritional value and stress adaptation. Accessions including teosintes, maize inbred lines and coix were investigated for kernel protein and its association with DNA markers. The proposed investigation assumed that wild accessions had different genic/allelic content and consequently expression profile than modern maize because of the domestication syndrome and bottleneck effects. Total protein content in hard stony fruit case teosinte accessions were assessed from kernels with and without seed coats, while protein content from coix and maize lines was evaluated from kernels only. The accessions were also subjected to molecular profiling using 84 SSR markers, and obtained genotypic data were used for population structure and association analysis. The results emphasize that teosintes have higher protein content (18.5% to 26.29%), followed by coix (18.26%), and the least among maize lines (9% to 11%). Among teosintes, without-seed-coat samples had 3–6% higher protein content than with-seed-coat samples. When compared to other teosinte species, Z. mays subsp. mexicana accessions showed higher protein content, ranging from 18.62% to 26.29%. All evaluated accessions were divided into four subpopulations with K = 4, and seven significant (p < 0.01) marker–trait associations were seen with umc1294, umc1171, phi091, umc2182 and bnlg292 markers, which are distributed across chromosomes 4, 5, 7, 8 and 9, respectively. We have observed that the wild relatives carry protein content-enhancing alleles and can be used as productive donor parents in pre-breeding efforts to increase the protein content of maize.
Pre-harvest sprouting (PHS) is a condition triggered by environmental factors, particularly prevalent in humid conditions, leading to substantial yield losses in black gram. While the potential for genotypic PHS tolerance exists, it has not been thoroughly assessed in black gram. Hence, the present study aimed to delve into the genetic variation for PHS tolerance in diverse black gram (Vigna mungo (L.) Hepper) germplasm, and also to comprehend the impact of various physical and physiological traits on PHS. A diverse set of 112 black gram accessions collected across the phytogeographical zones of India were examined for their seed and pod characteristics. Water absorption by pods and seeds and fresh-seed germination was calculated by following the standard procedure given by the International Seed Testing Association. The alpha-amylase activity was measured on dry seeds (0 h), 24 h, 48 h, and 72 h after germination of each accession, using a UV-VIS spectrophotometer, and hard-seededness was measured using a texture analyzer machine. The results showed a wide range in PHS tolerance and FSG, and 13 accessions were found to be PHS-tolerant (PHS value < 10%). An indicator of PHS, seed germination in a pod, ranged from 2.75% in IC485641 (highly tolerant to PHS) to 95.85% in IC530501 (highly susceptible to PHS). Correlation and multivariate analysis revealed that PHS was positively correlated with water imbibition by pod and seed, fresh-seed germination and alpha-amylase activity. PHS-tolerant accessions showed a slow increase in alpha-amylase activity, in contrast to PHS-susceptible accessions. The utilization of alpha-amylase activity as a biochemical marker has the potential for evaluating PHS tolerance across various black gram accessions. The identified PHS-tolerant accessions can be used as donors in crop improvement programs aimed at developing PHS-tolerant black gram varieties.