Genetic basis of plant regeneration in hexaploid triticale

Summary Glume cultures of monogenic recessive mutants PGTSLS1 (Plant Genetics triticale selection large spike 1) and PGTSLG2 (Plant Genetics triticale selection long grain 2) were employed along with their parent PGTS control (Plant Genetics triticale selection control) and their F1 and F2 progenies, to determine the genetic basis of plant regeneration in hexaploid triticale. The mutant PGTSLS1 exhibited greater efficiency of plant regeneration (22.4%) followed by PGTS control (7.6%) and PGTSLG2 did not exhibit any regeneration. All the three F1's exhibited plant regeneration frequency on par with that of control (6.9–7.3%), suggesting dominant nature of control genotype over the isogenic mutants. The F2 results suggested that genetic control over the high frequency regeneration of PGTSLS1 was monogenic recessive in nature, and genetic control over the recalcitrant nature of PGTSLG2 also was monogenic recessive. The F2 of the cross PGTSLS1 × PGTSLG2 segregated into four classes. Of the 114 F2 plants, 19 showed no regeneration, 70 of them exhibited 6–8% regeneration, 20 of them 19–24% regeneration, and 5 of them exhibited highest frequency regeneration (57–60%). These observations suggest dihybrid segregation for regeneration. The highest frequency of plant regeneration (57–60%) exhibited by 5 F2 plants may be due to the interaction of non-allelic genes in recessive condition. These results clearly demonstrate the association of at least two genes with plant regeneration in hexaploid triticale.     

Simple sequence repeats as useful resources to study transcribed genes of cotton

Microsatellites or Simple Sequence Repeats(SSRs) are informative molecular genetic markers in many crop species. SSRs are PCR-based, highly polymorphic, abundant, widely distributed throughout the genome and inherited in a co-dominant manner in most cases. Here we describe the presence of SSRs in cDNAs of cotton. Thirty one SSR primer pairs of 220 (∼14%) tested led to PCR amplification of discrete fragments using cotton leaf cDNA as template. Sequence analysis showed 25% of 24randomly selected cDNA clones amplified with different SSR primer pairs contained repeat motifs. We further showed that sequences from the SSR-containing cDNAs were conserved across G. barbadense and G. hirsutum, revealing the importance of the SSR markers for comparative mapping of transcribed genes. Data mining for plant SSR-ESTs from the publicly available databases identified SSRs motifs in many plant species,including cotton, in a range of 1.1 to4.8% of the submitted ESTs for a given species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Agrobacterium-mediated genetic transformation and production of semilooper resistant transgenic castor (Ricinus communis L.)

Summary Semilooper resistant transgenic castor plants were produced through Agrobacterium-mediated genetic transformation method. Two castor cultivars, Jyothi and VP1 were transformed using the super-binary vector pTOK233 carrying gus A and hpt genes. Putative transformants were regenerated following selection on the hygromycin containing medium. GUS positive primary transformants, when subjected to Southern analysis, revealed stable integration of gus A into their genomes. In the T₁ generation, a monogenic segregation ratio of 3 GUS positive: 1 GUS negative plants was observed. Furthermore, transformation experiments were carried out with the Agrobacterium pSB111 super-binary vector carrying a synthetic delta endotoxin gene cryIAb and the herbicide resistance gene bar both driven by cauliflower mosaic virus 35S promoter. Putative transformants were regenerated through selection on the phosphinothricin containing medium and Basta tolerant transformants were subjected to molecular analysis. PCR analysis revealed the presence of both bar and cryIAb genes in the Basta tolerant primary transformants. Southern analysis of PCR positive plants with cryIAb probe showed a 3 Kb band upon HindIII digestion and a > 6 Kb band with BamHI digestion, thus suggesting stable integration of cryIAb intact expression cassette and independent nature of the transformants. The primary transformants subjected to ELISA disclosed varied levels of Cry protein. These transgenics expressing cryIAb – when bioassayed against freshly hatched semilooper larvae – induced substantial (> 88%) insect mortality. Southern analysis of 2T₁ plants revealed the presence of cryIAb gene, indicating stable inheritance of the transgene into the next generation. In T₁, all the Southern-positive plants for cryIAb invariably exhibited tolerance to Basta, denoting co-segregation of both bar and cryIAb genes. Transgenics, expressing cryIAb exhibited ample resistance against the castor semilooper.     

Screening sorghum genotypes for salinity tolerant biomass production

Genetic improvement of salt tolerance is of high importance due to the extent and the constant increase in salt affected areas. Sorghum [Sorghum bicolor (L.) Moench] has been considered relatively more salt tolerant than maize and has the potential as a grain and fodder crop for salt affected areas. One hundred sorghum genotypes were screened for salinity tolerance in pots containing Alfisol and initially irrigated with a 250-mM NaCl solution in a randomized block design with three replications. Subsequently 46 selected genotypes were assessed in a second trial to confirm their responses to salinity. Substantial variation in shoot biomass ratio was identified among the genotypes. The performance of genotypes was consistent across experiments. Seven salinity tolerant and ten salinity sensitive genotypes are reported. Relative shoot lengths of seedlings were genetically correlated to the shoot biomass ratios at all stages of sampling though the relationships were not close enough to use the trait as a selection criterion. In general, the whole-plant tolerance to salinity resulted in reduced shoot Na⁺ concentration. The K⁺/Na⁺ and Ca²⁺/Na⁺ ratios were also positively related to tolerance but with a lesser r ². Therefore, it is concluded that genotypic diversity exists for salt tolerance biomass production and that Na⁺ exclusion from the shoot may be a major mechanism involved in that tolerance.     

Identification of quantitative trait loci associated with resistance to foliar diseases in sorghum [<Emphasis Type="Italic">Sorghum bicolor</Emphasis> (L.) Moench]

Forage sorghum cultivars grown in India are susceptible to various foliar diseases, of which anthracnose, rust, zonate leaf spot, drechslera leaf blight and target leaf spot cause severe damage. We report here the quantitative trait loci (QTLs) conferring resistance to these foliar diseases. QTL analysis was undertaken using 168 F₇ recombinant inbred lines (RILs) of a cross between a female parental line 296B (resistant) and a germplasm accession IS18551 (susceptible). RILs and parents were evaluated in replicated field trials in two environments. A total of twelve QTLs for five foliar diseases on three sorghum linkage groups (SBI-03, SBI-04 and SBI-06) were detected, accounting for 6.9–44.9% phenotypic variance. The morphological marker Plant color (Plcor) was associated with most of the QTL across years and locations. The QTL information generated in this study will aid in the transfer of foliar disease resistance into elite susceptible sorghum breeding lines through marker-assisted selection.     

Morph-physiological responses of cotton interspecific chromosome substitution lines to low temperature and drought stresses

Limited knowledge about genetic and physiological traits associated with drought and low temperature stresses and narrow genetic diversity in Upland cotton (Gossypium hirsutum L.) are serious impediments in its genetic improvement. The objectives of this research were to determine the genetic and physiological traits associated with drought and low temperature effects and to identify chromosomal effects on these traits using chromosome substitution (CS) lines from three alien species of Gossypium, G. barbadense, G. tomentosum, and G. mustelinum, respectively. Two experiments were conducted to study low temperature and drought stress effects during seedling emergence and early growth stages in 21 cotton CS-lines with parent, Texas Marker (TM)-1. In Experiment I, plants were grown at optimum (30/22 °C) and low (22/14 °C) temperature conditions under optimum water and nutrient conditions. In Experiment II, plants were grown at optimum water (soil moisture content of 0.167 m³ m^?3) and in drought (soil moisture content 0.105 m³ m^?3) conditions under optimum temperature conditions. Above- and below-ground growth traits including several root traits of the CS lines were assessed at 25 days after sowing. The findings suggest which substituted chromosome or chromosome segment from the alien species likely harbors one or more genes for higher and lower tolerance to low temperature, respectively. CS-T04 and CSB08sh showed higher and lower tolerance to low temperature, respectively and CS-T04 and CS-B22sh showed higher and lower tolerance, respectively, to drought. CS lines are valuable analytical tool and useful genetic resources for targeted exploitation of beneficial genes for drought and low temperature stresses in Upland cotton.     

Pigeonpea improvement: An amalgam of breeding and genomic research

In the past five decades, constant research has been directed towards yield improvement in pigeonpea resulting in the deployment of several commercially acceptable cultivars in India. Though, the genesis of hybrid technology, the biggest breakthrough, enigma of stagnant productivity still remains unsolved. To sort this productivity disparity, genomic research along with conventional breeding was successfully initiated at ICRISAT. It endowed ample genomic resource providing insight in the pigeonpea genome combating production constraints in a precise and speedy manner. The availability of the draft genome sequence with a large‐scale marker resource, oriented the research towards trait mapping for flowering time, determinacy, fertility restoration, yield attributing traits and photo‐insensitivity. Defined core and mini‐core collection, still eased the pigeonpea breeding being accessible for existing genetic diversity and developing stress resistance. Modern genomic tools like next‐generation sequencing, genome‐wide selection helping in the appraisal of selection efficiency is leading towards next‐generation breeding, an awaited milestone in pigeonpea genetic enhancement. This paper emphasizes the ongoing genetic improvement in pigeonpea with an amalgam of conventional breeding as well as genomic research.     

Characterization of the A4 cytoplasmic male-sterile lines of sorghum using RFLP of mtDNA

Three sorghum cytoplasmic male sterile lines CSV4 A(V), CSV4 A(G₁) and CSV4 A(M), grouped as A₄, were compared with a milo (A₁) and two other non-milo (A₂ and A₃) cytoplasms for their RFLP patterns of mitochondrial DNA (mtDNA). A 9.7 kb clone from pearl millet mtDNA discriminated each of the three A₄ entries whereas other maize and pearl millet mtDNA clones used could not distinguish this group completely. The molecular differences within the A₄ cytoplasmic group offer some explanation for the inconsistency in the fertility restoration behaviour of these A₄ lines obtained with a definite set of testers in the field. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification and characterization of EST-SSRs in finger millet (<Emphasis Type="Italic">Eleusine coracana</Emphasis> (L.) Gaertn.)

In recent years, microsatellites have become the most used markers for studying population genetic diversity. The increased availability of the DNA sequences has given the possibility to develop EST-derived SSR markers. A total of 1,927 ESTs of Eleusine coracana available in the NCBI database were mined for SSRs. Di-nucleotides are the most occurring motifs accounting for more than 50% of the repeats, of which GA was the most abundant motif and tetra-nucleotides are the least occurring motifs. Of the 132 markers identified, 30 primer pairs based were synthesized. SSR markers were used for variety discrimination and genetic assessment in 15 finger millet accessions; 20 primers showed polymorphism and 13 primers were identified as having a PIC value above 0.5. On the basis of the distribution of these polymorphic alleles, the 15 accessions were classified into two groups. This study has demonstrated the potential of EST-derived SSR primer pairs in finger millet. These primers will serve as valuable source for further breeding programs.