Wajad Nazeer

Introgression of cotton leaf curl virus-resistant genes from Asiatic cotton (Gossypium arboreum) into upland cotton (G. hirsutum).

Cotton is under the constant threat of leaf curl virus, which is a major constraint for successful production of cotton in the Pakistan. A total of 3338 cotton genotypes belonging to different research stations were screened, but none were found to be resistant against the Burewala strain of cotton leaf curl virus (CLCuV). We explored the possibility of transferring virus-resistant genes from Gossypium arboreum (2n = 26) into G. hirsutum (2n = 52) through conventional breeding techniques. Hybridization was done manually between an artificial autotetraploid of G. arboreum and an allotetraploid G. hirsutum, under field conditions. Boll shedding was controlled by application of exogenous hormones, 50 mg/L gibberellic acid and 100 mg/L naphthalene acetic acid. Percentage pollen viability in F(1) hybrids was 1.90% in 2(G. arboreum) x G. hirsutum and 2.38% in G. hirsutum x G. arboreum. Cytological studies of young buds taken from the F(1) hybrids confirmed that they all were sterile. Resistance against CLCuV in the F(1) hybrids was assessed through grafting, using the hybrid plant as the scion; the stock was a virus susceptible cotton plant, tested under field and greenhouse conditions. All F(1) cotton hybrids showed resistance against CLCuV, indicating that it is possible to transfer resistant genes from the autotetraploid of the diploid donor specie G. arboreum into allotetraploid G. hirsutum through conventional breeding, and durable resistance against CLCuV can then be deployed in the field.

A New Synthetic Amphiploid (AADDAA) between Gossypium hirsutum and G. arboreum Lays the Foundation for Transferring Resistances to Verticillium and Drought.

Gossypium arboreum, a cultivated cotton species (2n = 26, AA) native to Asia, possesses invaluable characteristics unavailable in the tetraploid cultivated cotton gene pool, such as resistance to pests and diseases and tolerance to abiotic stresses. However, it is quite difficult to transfer favorable traits into Upland cotton through conventional methods due to the cross-incompatibility of G. hirsutum (2n = 52, AADD) and G. arboreum. Here, we improved an embryo rescue technique to overcome the cross-incompatibility between these two parents for transferring favorable genes from G. arboreum into G. hirsutum. Our results indicate that MSB2K supplemented with 0.5 mgl-1 kinetin and 250 mg-1 casein hydrolysate is an efficient initial medium for rescuing early (3 d after pollination) hybrid embryos. Eight putative hybrids were successfully obtained, which were further verified and characterized by cytology, molecular markers and morphological analysis. The putative hybrids were subsequently treated with different concentrations of colchicine solution to double their chromosomes. The results demonstrate that four putative hybrid plants were successfully chromosome-doubled by treatment with 0.1% colchicine for 24 h and become amphiploid, which were confirmed by cytological observation, self-fertilization and backcrossing. Preliminary assessments of resistance at seedling stage indicate that the synthetic amphiploid showed highly resistant to Verticillium and drought. The synthetic amphiploid between G. hirsutum × G. arboreum would lay the foundation for developing G. arboreum-introgressed lines with the uniform genetic background of G. hirsutum acc TM-1, which would greatly enhance and simplify the mining, isolation, characterization, cloning and use of G. arboreum-specific desirable genes in future cotton breeding programs.

Diallel analysis to study the genetic makeup of spike and yield contributing traits in wheat (Triticum aestivum L.)

Five wheat genotypes were crossed in complete diallel fashion for gene action studies of spike length, spikelets per spike, grains per spike, grain weight per spike and grain yield per plant. Analysis of variance reveals significant differences for all the traits. The significant deviation of joint regression value “b” from zero and non significance of “t” square value, suggested the absence of epistasis for all traits which in turn attested fitness of the data for simple additive dominance model. Additive genetic component (D) proved to be significant for all traits under consideration except grain weight spike per spike, but dominant component H 1 and H 2 was higher in magnitude than additive (D) for spike length and grain yield per plant, illustrating the prevalence of dominance genetic effects. The additive genetic component was more imperative for genetic manipulation of number of spikelets per spike and number of grains per spike. Dominant genes at most of the loci were in excess than recessive genes for spike length and grain yield per plant which was firmly supported by the positive value of F and further strengthened by the value of (4DH 1 ) 0.5 +F/ (4DH 1 ) 0.5 -F which was greater than unity for these two traits. The component that was strongly influenced by the environment was spikelets per spike. High magnitude of narrow sense heritability (h 2 n.s ) was noticed for spikelets per spike (79%), and grains per spike (88%) thus illustrated fixable and additive heritable variation for these traits. The operation of over dominance was observed for spike length, grain weight per spike and grain yield per plant, whereas, spikelets per spike and grains per spike were under additive type of gene action with partial dominance, implying that spikelets per spike and grains per spike can significantly be improve by pursuing pedigree method while heterosis can be exploited for spike length, grain weight per spike and grain yield per plant. Key words: Wheat, gene action, genetic parameters, heritability, yield.

Introgression of genes for cotton leaf curl virus resistance and increased fiber strength from Gossypium stocksii into upland cotton (G. hirsutum).

Cotton leaf curl virus disease is a major hurdle for successful cotton production in Pakistan. There has been considerable economic loss due to this disease during the last decade. It would be desirable to have cotton varieties resistant to this disease. We explored the possibility of transferring virus resistant genes from the wild species Gossypium stocksii into MNH-786, a cultivar of G. hirsutum. Hybridization was done under field condition at the Cotton Research Station, Multan, during 2010-11. Boll shedding was controlled by application of exogenous hormones. F1 seeds were treated with 0.03% colchicine solution for 6 h and germinated. Cytological observations at peak squaring/flowering stage showed that these plants were hexaploid, having 2n = 6x = 78 chromosomes. The F1 plants showed intermediate expression for leaf size, leaf area, petiole length, bracteole number and size, bracteole area, bracteole dentation, flower size, pedicel size, and petal number and size. Moreover it possessed high fiber strength of 54.4 g/tex, which is 54% greater than that of the check variety, i.e. MNH-786 (G. hirsutum). The F1 population did not show any symptom of CLCuVD in the field, tested by grafting with CLCuVD susceptible rootstock (var. S12). We conclude that it is possible to transfer CLCuVD resistance and high fiber strength from G. stocksii to G. hirsutum.

Evaluation of cotton leaf curl virus resistance in BC1, BC2, and BC3 progenies from an interspecific cross between Gossypium arboreum and Gossypium hirsutum.

Cotton leaf curl virus disease (CLCuD) is an important constraint to cotton production. The resistance of G. arboreum to this devastating disease is well documented. In the present investigation, we explored the possibility of transferring genes for resistance to CLCuD from G. arboreum (2n = 26) cv 15-Mollisoni into G. hirsutum (2n = 52) cv CRSM-38 through conventional breeding. We investigated the cytology of the BC1 to BC3 progenies of direct and reciprocal crosses of G. arboreum and G. hirsutum and evaluated their resistance to CLCuD. The F1 progenies were completely resistant to this disease, while a decrease in resistance was observed in all backcross generations. As backcrossing progressed, the disease incidence increased in BC1 (1.7–2.0%), BC2 (1.8–4.0%), and BC3 (4.2–7.0%). However, the disease incidence was much lower than that of the check variety CIM-496, with a CLCuD incidence of 96%. Additionally, the disease incidence percentage was lower in the direct cross 2(G. arboreum)×G. hirsutum than in that of G. hirsutum×G. arboreum. Phenotypic resemblance of BC1 ∼BC3 progenies to G. arboreum confirmed the success of cross between the two species. Cytological studies of CLCuD-resistant plants revealed that the frequency of univalents and multivalents was high in BC1, with sterile or partially fertile plants, but low in BC2 (in both combinations), with shy bearing plants. In BC3, most of the plants exhibited normal bearing ability due to the high frequency of chromosome associations (bivalents). The assessment of CLCuD through grafting showed that the BC1 to BC3 progenies were highly resistant to this disease. Thus, this study successfully demonstrates the possibility of introgressing CLCuD resistance genes from G. arboreum to G. hirsutum.

Determination of genetic variation among chickpea genotypes and their F1 crosses with RAPD markers.

The intensity of genetic diversity amongst chickpea genotypes and their crosses is unknown. The current study investigated the genetic diversity of chickpea genotypes and their F₁ crosses by using random amplified polymorphic DNA (RAPD) markers. We assessed the variation among six chickpea genotypes and 15 F₁ crosses with the RAPD markers. The six parents and their 21 hybrids were carefully studied based on the presence or absence of bands. The level of polymorphism varied with different primers. Out 28 primers used, 21 amplified the genomic DNA in all the varieties, 15 generated polymorphic bands among all the varieties, and six produced similar banding patterns.