Recent advances in molecular marker techniques: Insight into QTL mapping,GWAS and genomic selection in plants

Recent advances in sequencing technology have brought several novel platforms for marker development and subsequent genotyping. The high-throughput and cost effective marker techniques have changed the entire scenario of marker applications. The huge genotypic data obtained with next generation sequencing (NGS) also demands analytical tools, statistical advances, and comprehensive understanding to cope with breeding applications. In the present review, we discussed different available marker techniques, their strengths, and limitations. Emphasis was given on software tools, analytical pipelines, workbenches, and online resources available for marker development. Comparison of SNP genotyping involving complexity reduction techniques like GBS, RRL, RAD, and array-based platforms were presented in a view to describe suitability for specific purposes. We found that genotyping by whole genome re-sequencing has great potential, and could be a routine application in the near future with continuously decreasing cost of sequencing. Microsatellites, still a valuable option for breeders, have also advanced with NGS. Here a catalogue of tools for microsatellite evaluation in short sequence reads was provided. The most common applications of molecular marker like QTL mapping, genome-wide association mapping (GWAS), and genomic selection were highlighted. The present review will be helpful for the effective utilization of available resources and for the planning of crop improvement programs employing molecular marker techniques.     

Widening the gene pool of cultivated lentils through introgression of alien chromatin from wild Lens subspecies

Wild Lens species/subspecies are a potential source for increasing genetic diversity in cultivated lentil. Four intraspecific crosses were attempted between cultivated and wild lentils. Viable hybrids were produced between L. culinaris ssp. culinaris × L. culinaris ssp. orientalis and L. culinaris ssp. culinaris × L. culinaris ssp. odomensis. Normal meiosis and pollen fertility were observed in the first set of crosses, whereas chromosomal abnormalities and reduced pollen fertility were observed in the second set of crosses. These crosses were also studied for some quantitative traits. The range, mean and coefficient of variation were calculated in parents, F₁, F₂ and BC₁ generations to determine the extent of variability generated in the cultivated lentil through introgression of genes from wild lentil. The cultivated lentil × L. culinaris ssp. orientalis crosses showed substantially higher variability for all the traits than crosses involving cultivated lentil ×L. culinaris ssp. odomensis. The results of the present study indicated that these wild subspecies can be exploited for breeding purposes and their variation can easily be utilized to widen the genetic base of the cultivated lentil.     

Male-sterility in the grain amaranth (Amaranthus hypochondriacus ex-Nepal) variety Jumla

Summary Twenty naturally occurring male-sterile plants were recovered from a normal population of the grain amaranth, variety Jumla. The identification of the male-sterile plants is possible during anthesis and after flowering. The male-sterility in this variety is conditioned by a single recessive nuclear gene ms.This research was supported by Grant No. AMA-KE-4-83-22 (CRG GRANT) from the National Academy of Science, USA. Reprint requests to V.K. Gupta.     

Genetic Analysis of Resistance to Xanthomonas campestris pv. oryzae (Ishiyama) Dye in a Rice Breeding Line ARC 10464

The genetics of resistance to three Indian pathotypes of Xanthomonas campestris pv. oryzae, namely, IX01, IX08 and IX09, was studied in a landrace of Indica rice ‘ARC 10464’. Resistance to each of the two pathotypes IX01 and IX09 was governed by two independently-inherited dominant genes while a single dominant gene was operative against patho-type 1X08. The joint segregation tests conducted on F₂ plant progenies (F₃ families) using pathotypes IX01, IX08 and IX09 suggested that the gene/(s) effective against each of the pathotypes are different.     

A new gene for resistance in rice to Asian rice gall midge (Orseolia oryzae Wood Mason) biotype 1 population at Raipur, India

The Asian rice gall midge, Orseolia oryzae Wood Mason (Diptera: Cecidomyiidae), is a major pest of rice in several South and South East Asian countries. The maggots feed internally on the growing tips of the tillers and transform them into tubular galls, onion leaf-like structures called ‘silver shoots’ resulting into severe yield loss to the rice crop. We studied the mode of inheritance and allelic relationships of the resistance genes involved in resistant donor Line 9, a sib of a susceptible cultivar ‘Madhuri’. The segregation behaviour of F₁, F₂ and F₃ populations of the cross between Line 9 and susceptible cultivar MW10 confirmed the presence of a single dominant gene for resistance. Tests of allelism with all the known genes giving resistance to this population indicated that Line 9 possessed a new gene which was designated Gm 9 This revised version was published online in July 2006 with corrections to the Cover Date.     

Inheritance of growth vigour and its association with other characters in chickpea

Growth vigour plays an important role in the establishment of a normal crop. The F₂ population of a cross between high‐ and low‐growth vigour varieties of chickpea segregated into 15 high : 1 low growth vigour. The results for recombinant inbred lines and BC1P₂ showed a good fit to the expected 3 : 1 ratio. The results indicated that growth vigour is controlled by two genes with duplicate dominant epistasis. No gene has so far been identified for growth vigour in chickpea. Correlation between growth vigour and other characters showed that high growth vigour had significant negative correlation with days to first flower, days to 50% flowering, days to first pod and days to maturity.     

Effect of Water Stress on Physiological Attributes and their Relationship with Growth and Yield of Wheat Cultivars at Different Stages

The effects of water stress on physiological attributes of drought‐sensitive (Kalyansona) and drought‐tolerant (C‐306) wheat cultivars were studied in a pot experiment. Water stress was imposed by withholding irrigation at boot and anthesis stages. Leaf water potential, leaf osmotic potential and leaf turgor potential (measured with pressure chamber and osmometer), as well as leaf diffusive resistance, leaf transpiration rate and leaf‐to‐air transpiration gradient (measured with a steady‐state porometer) were measured diurnally. Growth and yield parameters were recorded after harvesting of the crop. Triplicate data were analysed using a completely randomized design and correlations amongst these parameters were computed. Water stress was found to reduce diurnal leaf water potential and leaf osmotic potential in both the genotypes but leaf osmotic potential was significantly higher in the drought‐tolerant cultivar C‐306 than in the drought‐sensitive cultivar Kalyansona. Positive turgor was recorded in both the genotypes under water stress and non‐stress conditions. Water‐stressed plants showed significantly lower turgor potential than control plants. In diurnal observations, water‐stressed plants exhibited significantly higher leaf diffusive resistance in both genotypes at both stages. The diffusive resistance of C‐306 was predominantly higher than that of Kalyansona. Water stress decreased leaf transpiration rate at both stages but the reduction was higher at the anthesis stage. The leaf‐to‐air temperature gradient was much higher in C‐306 than in Kalyansona at the boot stage but at the anthesis stage genotypic variation was non‐significant. The capacity to maintain cooler foliage was lower at the anthesis stage than at the boot stage in both the cultivars. Shoot dry weight, number of grains, test weight, grain yield, biological yield and harvest index decreased to a greater extent when water stress was imposed at the anthesis stage, while imposition of water stress at the boot stage caused a greater reduction in plant height and number of tillers. Similarly, water stress caused a smaller reduction in growth, yield and yield attributes in C‐306 than in Kalyansona. In general, the correlation coefficient of grain and biological yield with water potential and its components was positive and highly significant. Similarly, turgor potential was also correlated positively and significantly with grain yield at both the stages, but with biological yield it was significant only at the anthesis stage. A negative and significant correlation was obtained for diffusive resistance and leaf‐to‐air temperature gradient with grain yield at the boot and anthesis stages. The rate of transpiration was also positively and significantly correlated to grain and biological yields at both the stages. Amongst the yield attributes, number of leaves and number of tillers were positively correlated at the anthesis stage, whereas leaf area and shoot dry weight were significantly correlated with grain and biological yields at both the stages.     

Genes Lr48 and Lr49 for hypersensitive adult plant leaf rust resistance in wheat (Triticum aestivum L.)

The genetic bases of leaf rust resistance in wheat (Triticum aestivum L.) line CSP44, selected from the Australian cultivar Condor, and Indian cultivar VL404, were studied. The reaction patterns of CSP44 and VL404 against Indian races 12, 77, 77-1, 77-2, 77-3, 77-4, 77-5 and 108 were different from reaction patterns shown by near-isogenic lines with known adult plant resistance (APR) genes, viz. Lr12, Lr13, Lr22b and Lr34. Although the reaction patterns of CSP44 and VL404 were similar to the near-isogenic line Tc+Lr22a, tests of allelism indicated absence of Lr22a in both CSP44 and VL404. On the basis of genetic studies, their resistances in field tests against race 77-5, the most virulent race from the Indian sub-continent, were each ascribed to two genes. One of the two genes in each wheat was identified to be the non-hypersensitive APR gene Lr34. The second APR genes in CSP44 and VL404 gave hypersensitive reaction types and were recessive and dominant, respectively. The gene in CSP44 was designated Lr48and the gene in VL404, Lr49. This revised version was published online in August 2006 with corrections to the Cover Date.     

SCAR marker tagged to the alien leaf rust resistance gene Lr19 uniquely marking the Agropyron elongatum-derived gene Lr24 in wheat: a revision

A sequence characterized amplified region (SCAR) marker tagged to an Agropyron elongatum‐derived leaf rust resistance (Lr) gene Lr19 was validated on 18 known alien Lr gener in near‐isogenic lines (NILs) in the variety ‘Thatcher’, along with three wheat cultivers carrying Lr24 and two carrying Lr19. The marker was expressed only in the Lr24 lines confirming that the marker tagged the geneLr24. The monomorphic expression of the SCAR marker in 10NIL pairs for Lr19 and Lr24 revealed that each NIL pair possessed the same gene, Lr24. The donor parents used in the NIL pairs for Lr19 (‘Sunstar*6/C80‐1′) and Lr24 (‘TR380‐14*7/3Ag#14′) amplified the same fragment. Nonsegregation for leaf rust in the F₂ population of the cross between the above donor parents confirmed the presence of the same gene in the two parents. Apparently, a genuine parent stock of ‘Sunstar*6/C80‐1’ was not involved in the development of the NIL pairs for Lr19 due to an improper maintence bredding protocol either at source or destination which went undetected in the absence of signs of virulence for either gene in the region.