Toward allotetraploid cotton genome assembly: integration of a high-density molecular genetic linkage map with DNA sequence information.

BACKGROUND: Cotton is the world's most important natural textile fiber and a significant oilseed crop. Decoding cotton genomes will provide the ultimate reference and resource for research and utilization of the species. Integration of high-density genetic maps with genomic sequence information will largely accelerate the process of whole-genome assembly in cotton. RESULTS: In this paper, we update a high-density interspecific genetic linkage map of allotetraploid cultivated cotton. An additional 1,167 marker loci have been added to our previously published map of 2,247 loci. Three new marker types, InDel (insertion-deletion) and SNP (single nucleotide polymorphism) developed from gene information, and REMAP (retrotransposon-microsatellite amplified polymorphism), were used to increase map density. The updated map consists of 3,414 loci in 26 linkage groups covering 3,667.62 cM with an average inter-locus distance of 1.08 cM. Furthermore, genome-wide sequence analysis was finished using 3,324 informative sequence-based markers and publicly-available Gossypium DNA sequence information. A total of 413,113 EST and 195 BAC sequences were physically anchored and clustered by 3,324 sequence-based markers. Of these, 14,243 ESTs and 188 BACs from different species of Gossypium were clustered and specifically anchored to the high-density genetic map. A total of 2,748 candidate unigenes from 2,111 ESTs clusters and 63 BACs were mined for functional annotation and classification. The 337 ESTs/genes related to fiber quality traits were integrated with 132 previously reported cotton fiber quality quantitative trait loci, which demonstrated the important roles in fiber quality of these genes. Higher-level sequence conservation between different cotton species and between the A- and D-subgenomes in tetraploid cotton was found, indicating a common evolutionary origin for orthologous and paralogous loci in Gossypium. CONCLUSION: This study will serve as a valuable genomic resource for tetraploid cotton genome assembly, for cloning genes related to superior agronomic traits, and for further comparative genomic analyses in Gossypium.

A cascade of recently discovered molecular mechanisms involved in abiotic stress tolerance of plants.

Today, agriculture is facing a tremendous threat from the climate change menace. As human survival is dependent on a constant supply of food from plants as the primary producers, we must aware of the underlying molecular mechanisms that plants have acquired as a result of molecular evolution to cope this rapidly changing environment. This understanding will help us in designing programs aimed at developing crop plant cultivars best suited to our needs of a sustainable agriculture. The field of systems biology is rapidly progressing, and new insight is coming out about the molecular mechanisms involved in abiotic stress tolerance. There is a cascade of changes in transcriptome, proteome, and metabolome of plants during these stress responses. We have tried to cover most pronounced recent developments in the field of "omics" related to abiotic stress tolerance of plants. These changes are very coordinated, and often there is crosstalk between different components of stress tolerance. The functions of various molecular entities are becoming more clear and being associated with more precise biological phenomenon.

Pistil drip following pollination: a simple in planta Agrobacterium-mediated transformation in cotton.

Transgenic cotton plants were developed by pistil drip inoculation in a solution containing Agrobacterium carrying a gene for resistance to the herbicide Basta (bar), 10% (w/v) sucrose, 0.05% (v/v) Silwet L-77 and 40 mg acetosyringone l(-1). Pistil drip during 17:00-19:00 on the first day of flowering resulted in 0.07-0.17% Basta-resistant plants/number of viable seeds generated, and stigma excision prior to pistil drip during this time period gave rise to a transformation efficiency of 0.46-0.93%, in contrast with 0.04-0.06% generated from pistil drip during 9:00-11:00 on the second day of flowering. PCR and Southern blot analysis confirmed the integration of the bar gene into the cotton genome, and a T1 and T2 generation herbicide resistance test consistently revealed expression and stable heritability of the bar gene in the two generations.

Molecular cloning and characterization of a cytosolic glutamine synthetase gene, a fiber strength-associated gene in cotton.

An expressed sequence tag encoding glutamine synthetase (GS) was identified by microarray-based hybridization using fiber mRNAs of allotetraploid Gossypium hirsutum, 7235, a super quality property germplasm line, and TM-1, a genetic standard in G. hirsutum. Northern-blot analysis verified transcript accumulation differences in 8 DPA fibers (including ovules) in the two varieties. The full-length cDNA encoding GS in 7235 was isolated and named GhGS. Sequence analysis revealed that the GhGS was similar to cytosolic GS. Southern-blot analysis showed that tetraploid cotton contained at least one copy of the A sub-genome and the D sub-genome. Genomic GhGS sequences were subsequently isolated from different varieties, TM-1, 7235 and two diploid progenitor cottons, G. herbaceum (A-genome) and G. raimondii (D-genome). Molecular mapping and single-marker analysis revealed that the GhGS was significantly correlated with fiber strength and was mapped to chromosome D7. Additionally, GS activities and total protein of the ovules and the fibers were assayed. The results showed a significantly higher GS activity in 7235 seeds compared to TM-1 seeds at 5 and 8 DPA. Also significant differences were found in total protein content and seed weight at 11 DPA. This suggested that GS promoted the seed-forming process by providing N. On the other hand, in fibers, GS activity and total protein assay indicated a lower total GS activity and longer fiber elongation period in 7235. These results suggest that the respective roles of the GS in ovules and fibers do not completely overlap.