Genome wide identification and functional characterization of the cotton C-repeat binding factor (CBF) under cold stress condition

Background Low temperature is a common biological abiotic stress in major cotton growing areas. Cold stress significantly affects the growth, yield and yield quality of cotton. Therefore, it is important to develop a more robust and cold stress tolerant cotton germplasms. In response to climate change and erratic conditions, plants have evolved various survival mechanisms, one of which induction of various stress responsive transcription factors, such as the C-repeat binding factors (CBFs), which have been found to enhance cold tolerance in various plants. Results In this study detailed evaluation of the cotton CBF has been carried out. A total of 29, 28, 25, 21, 30, 26 and 15 proteins encoded by the CBF genes were identified in Gossypium herbaceum, Gossypium arboreum, Gossypium thurberi, Gossypium raimondii, Gossypium turneri, Gossypium longicalyx and Gossypium australe, respectively. Phylogenetic evaluation revealed that the proteins were grouped into seven clades, with clade 1 and 6 being the largest. Moreover, majority of the proteins encoded by the genes were predicted to be located within the nucleus, while some were distributed in other parts of the cell. Based on the transcriptome and RT-qPCR analysis, Gthu17439 (GthCBF4) was highly upregulated under cold stress, and was further validated through forward genetics. The Gthu17439 (GthCBF4) overexpressed plants showed a significantly tolerance to cold stress, with higher germination rate, higher root growth and high induction levels of stress responsive genes. The over-expressed plants exhibited low level of oxidative damage, due to significant reduction in the H2O2 production. Conclusion The results showed that the Gthu17439 (GthCBF4) could be playing a significant role in enhancing cold stress tolerance in cotton and can be further exploited in developing cotton germplasm with an improved cold-stress tolerance

Comparative transcriptome, physiological and biochemical analyses reveal response mechanism mediated by CBF4 and ICE2 in enhancing cold stress tolerance in Gossypium thurberi

Low temperature is one of the key environmental stresses that impair plant growth and significantly restricts the productivity and spatial distribution of crop plants. Gossypium thurberi, a wild diploid cotton species, has adapted to a wide range of temperatures and exhibits a better tolerance to chilling stress. Here, we compared phenotypes and physiochemical changes in G. thurberi under cold stress and found this species indeed showed better cold tolerance. Therefore, to understand the molecular mechanisms of the cold tolerance in G. thurberi, we compared transcription changes in leaves of G. thurberi under cold stress by high-throughput transcriptome sequencing. In total, 35 617 unigenes were identified in the whole-genome transcription profile, and 4226 differentially expressed genes (DEGs) were discovered in the leaves upon cold treatment. Gene Ontology (GO) classification analyses showed that the majority of DEGs belonged to categories of signal transduction, transcription factors (TFs) and carbohydrate transport and metabolism. The expression of several cold-responsive genes such as ICE1, CBF4, RAP2-7 and abscisic acid (ABA) biosynthesis genes involved in different signalling pathways were induced after G. thurberi seedlings were exposed to cold stress. Furthermore, cold sensitivity was increased in CBF4 and ICE2 virus-induced gene silencing (VIGS) plants, and high level of malondialdehyde (MDA) showed that the CBF4 and ICE2 silenced plants were under oxidative stress compared to their wild types, which relatively had higher levels of antioxidant enzyme activity, as evident by high levels of proline and superoxide dismutase (SOD) content. In conclusion, our findings reveal a new regulatory network of cold stress response in G. thurberi and broaden our understanding of the cold tolerance mechanism in cotton, which might accelerate functional genomics studies and genetic improvement for cold stress tolerance in cultivated cotton.

Genetic map construction and functional characterization of genes within the segregation distortion regions (SDRs) in the F2:3 generation derived from wild cotton species of the D genome

Segregation distortion (SD) is a phenomenon common among stable or segregating populations, and the principle behind it is still an issue that puzzles many researchers. An F2:3 generations developed from the wild cotton species of the D genomes was applied to investigate the possible plant transcription factors within the segregation distortion regions (SDRs). We constructed a consensus map between two maps in the D genome, map A; Gossypium klotzschianum and Gossypium davidsonii and Map B; Gossypium thurberi and Gossypium trilobum. The two maps were developed from 188 F2:3 populations for each map, a total of 1492 markers, were linked to the 13 linkage groups. The consensus linkage map size was 1467.445 cM with an average marker distance of 1.0370cM. Chr02 had the highest percentage of segregation distortion with 58.621% followed by Chr07 with 47.887%. A total of 6,038 genes were mined within the segregation distortion regions (SDR region) of Chr02 and Chr07 with 2,308 gene in Chr02 and 3,730 genes in Chr07, we obtained a total of 1,117 domains within the SDR with a total of 622 domains shared between the two chromosomes, the first 9 domains all belonged to the plant resistance genes (R genes), the largest domain was PF00069 with a total of 188 genes. A total of 287 miRNAs were found to target the various genes, such as gr-miR398, gra-miR5207, miR164a, miR164b, miR164c among others which have been found to target top-ranked stress-responsive transcription factors such as NAC genes. Moreover, the genes were found to be regulated by various stress responsive cis-regulatory elements. RNA profiling showed that higher numbers of genes were highly upregulated in abiotic and different fiber development stages. The result shows that the SDR regions could be playing an important role in the evolution of significant genes in plants.

Genetic map construction and functional characterization of genes within the segregation distortion regions (SDRs) in the F2:3 generation derived from wild cotton species of the D genome

Segregation distortion (SD) is a phenomenon common among stable or segregating populations, and the principle behind it is still an issue that puzzles many researchers. An F2:3 generations developed from the wild cotton species of the D genomes was applied to investigate the possible plant transcription factors within the segregation distortion regions (SDRs). We constructed a consensus map between two maps in the D genome, map A; Gossypium klotzschianum and Gossypium davidsonii and Map B; Gossypium thurberi and Gossypium trilobum. The two maps were developed from 188 F2:3 populations for each map, a total of 1492 markers, were linked to the 13 linkage groups. The consensus linkage map size was 1467.445 cM with an average marker distance of 1.0370cM. Chr02 had the highest percentage of segregation distortion with 58.621% followed by Chr07 with 47.887%. A total of 6,038 genes were mined within the segregation distortion regions (SDR region) of Chr02 and Chr07 with 2,308 gene in Chr02 and 3,730 genes in Chr07, we obtained a total of 1,117 domains within the SDR with a total of 622 domains shared between the two chromosomes, the first 9 domains all belonged to the plant resistance genes (R genes), the largest domain was PF00069 with a total of 188 genes. A total of 287 miRNAs were found to target the various genes, such as gr-miR398, gra-miR5207, miR164a, miR164b, miR164c among others which have been found to target top-ranked stress-responsive transcription factors such as NAC genes. Moreover, the genes were found to be regulated by various stress responsive cis-regulatory elements. RNA profiling showed that higher numbers of genes were highly upregulated in abiotic and different fiber development stages. The result shows that the SDR regions could be playing an important role in the evolution of significant genes in plants.

Production of New Cotton Interspecific Hybrids with Enhanced Fiber Fineness

<p>To improve cotton fiber fineness, the (<em>Gossypium hirsutum</em> L. × <em>Gossypium longicalyx </em>Hutch. &amp; Lee)² allohexaploid and the [(<em>Gossypium hirsutum</em> L. × <em>Gossypium thurberi </em>Tod.)² × <em>G. longicalyx</em>] allotetraploid were backcrossed to <em>G. hirsutum</em> to produce introgressed genetic stocks. The ribbon width (RW) of 600 swelled fibers produced by the hybrids, their parents, and their backcross progeny were analyzed for each compared genotype using an optical microscope. The RWs varied between 6.41±2.15 µm for <em>G. longicalyx</em> to 17.45±2.98 µm for the <em>G. hirsutum</em> parent cultivar C2. Fibers produced by the trispecific hybrids and their progeny were finer than the bispecific hybrid material. For the introgressed stocks, the lowest RWs were observed for the trispecific hybrid (10.79±2.14 µm) and certain backcross progenies (between 11.98±1.27 µm to 12.71±1.61 µm). The allohexaploid RW was 13.58±1.41 µm. One of its tetraploid progeny produced approximately the same value (13.94±2.48 µm). These results show that <em>G. longicalyx </em>is a potential genetic stock for cotton fiber fineness improvement. The genetic stocks produced are valuable materials for improve the fineness of cotton fiber.</p>