Analysis of genetic diversity and population structure in upland cotton (Gossypium hirsutum L.) germplasm using simple sequence repeats.

Improvement of cotton fibre yield and quality is challenging due to the narrow genetic base of modern cotton cultivars, which emphasizes the great need to effectively explore the existing germplasm resources. With major objective to assess the genetic diversity and population structure at DNA level, 302 elite upland cotton germplasm accessions (253 Chinese and 49 different exotic origins), were genotyped using 198 simple sequence repeats (SSRs) markers. Each of the 198 markers differed greatly in its ability to detect variations in the panel of cotton germplasm. The SSRs amplified 897 alleles, of which 77.7% were polymorphic. The number of alleles varied from 2 to 12 (mean 4.53). Gene diversity ranged from 0.020 to 0.492 with a mean of 0.279. The polymorphic information content (PIC) values ranged from 0.371 to 0.019 (mean 0.225). Genetic distances in the whole cotton germplasm ranged from 0.451 to 0.052 (mean 0.270), demonstrating relatively wider genetic diversity range. Chinese-origin cotton germplasm showed the highest level of SSR polymorphisms (gene diversity=0.268, PIC=0.218), whereas American-origin revealed the highest mean genetic distance (0.274). Model-based Bayesian analysis clustered the whole cotton germplasm into three subpopulations, and the highest molecular variation ws revealed between subpopulations (4%, P<0.001). The SSRs revealed moderate level of genetic diversity at DNA level, identified three structured subpopulations, suggesting a potential use of these markers for genomewide association mapping studies and for identifying and conserving useful alleles in upland cotton germplasm.

QTL Mapping of Fiber Quality and Yield-Related Traits in an Intra-Specific Upland Cotton Using Genotype by Sequencing (GBS).

Fiber quality and yield improvement are crucial for cotton domestication and breeding. With the transformation in spinning techniques and multiplicity needs, the development of cotton fiber quality and yield is of great importance. A genetic map of 5178 Single Nucleotide Polymorphism (SNP) markers were generated using 277 F2:3 population, from an intra-specific cross between two upland cotton accessions, CCRI35 a high fiber quality as female and Nan Dan Ba Di Da Hua (NH), with good yield properties as male parent. The map spanned 4768.098 cM with an average distance of 0.92 cM. A total of 110 Quantitative Traits Loci (QTLs) were identified for 11 traits, but only 30 QTLs were consistent in at least two environments. The highest percentage of phenotypic variance explained by a single QTL was 15.45%. Two major cluster regions were found, cluster 1 (chromosome17-D03) and cluster 2 (chromosome26-D12). Five candidate genes were identified in the two QTL cluster regions. Based on GO functional annotation, all the genes were highly correlated with fiber development, with functions such as protein kinase and phosphorylation. The five genes were associated with various fiber traits as follows: Gh_D03G0889 linked to qFM-D03_cb, Gh_D12G0093, Gh_D12G0410, Gh_D12G0435 associated with qFS-D12_cb and Gh_D12G0969 linked to qFY-D12_cb. Further structural annotation and fine mapping is needed to determine the specific role played by the five identified genes in fiber quality and yield related pathway.

Gene mapping and transcriptome profiling of a practical photo-thermo-sensitive rice male sterile line with seedling-specific green-revertible albino leaf.

Abnormal environment weather can cause rice photoperiod-thermo-sensitive genic male sterile (PTGMS) lines fertile or partially fertile and thus cause the mixture of true hybrids with selfing seeds. Seedling-specific green-revertible albino leaf color mutant can be used to distinguish the real hybrids. Besides, it can also be used as an ideal material to research the development of chloroplast and biosynthesis of chlorophyll. The phenotype of leaf color mutants includes light green, yellowing, albino, green-revertible albino. Gene mutations affecting the synthesis and degradation of photosynthetic pigments, lycopene and heme, the differentiation and development of chloroplast, gibberellins (GAs) biosynthesis, can change the leaf color. We have created a PTGMS line with seedling-specific green-revertible albino leaf named W01S. The leaf phenotype, pollen sterility and fertility, agronomic traits, heredity, gene mapping and RNA-Seq of the differentially expressed genes between albino and green-revertible leaves were investigated. The results showed that W01S is a practical PTGMS line as Pei'ai 64S. The mutation of candidate gene Os03g0594100 (ent-isokaurene C2-hydroxylase-like) in W01S can be related to the biosynthesis of GAs, indole acetic acids, ethylene.

High-Density Linkage Map Construction and Mapping of Salt-Tolerant QTLs at Seedling Stage in Upland Cotton Using Genotyping by Sequencing (GBS).

Over 6% of agricultural land is affected by salinity. It is becoming obligatory to use saline soils, so growing salt-tolerant plants is a priority. To gain an understanding of the genetic basis of upland cotton tolerance to salinity at seedling stage, an intra-specific cross was developed from CCRI35, tolerant to salinity, as female with Nan Dan (NH), sensitive to salinity, as the male. A genetic map of 5178 SNP markers was developed from 277 F2:3 populations. The map spanned 4768.098 cM, with an average distance of 0.92 cM. A total of 66 QTLs for 10 traits related to salinity were detected in three environments (0, 110, and 150 mM salt treatment). Only 14 QTLs were consistent, accounting for 2.72% to 9.87% of phenotypic variation. Parental contributions were found to be in the ratio of 3:1, 10 QTLs from the sensitive and four QTLs from the resistant parent. Five QTLs were located in At and nine QTLs in the Dt sub-genome. Moreover, eight clusters were identified, in which 12 putative key genes were found to be related to salinity. The GBS-SNPs-based genetic map developed is the first high-density genetic map that has the potential to provide deeper insights into upland cotton salinity tolerance. The 12 key genes found in this study could be used for QTL fine mapping and cloning for further studies.

Identification and characterization of cotton genes involved in fuzz-fiber development.

Cotton fuzz fibers, like Arabidopsis trichomes, are elongated unicells. It is postulated that a transcriptional complex of GLABRA1 (GL1), GLABRA3 (GL3), and TRANSPARENT TESTAGLABRA1 (TTG1) might be in existence in Arabidopsis as evidenced by their physical interaction in yeast, and the complex regulates expression of GLABRA2 (GL2) controlling trichome cell differentiation; it is also assumed that TRIPTYCHON (TRY) and CAPRICE (CPC) counteract the complex formation in neighboring cells. Here, the homologs GaMYB23 (a homolog of GL1), GaDEL65 (a homolog of GL3), GaTTG1, GaCPC and GaTRY were identified in Gossypium arboreum. We show that GaMYB23 can bind to and activate the promoters of GaCPC, GaGL2 and GaTRY, and that GaMYB23, GaTRY and GaTTG1 could interact with GaDEL65 in yeast and in planta. In situ analysis showed that GaMYB23, GaGL2, GaDEL65, and GaTRY were predominantly expressed in fuzz fiber, but GaTRY proteins were primarily found in undeveloped epidermal cells. A G. arboreum fuzzless mutant with consistently high level GaMYB23 transcript has lost the detectable GaMYB23-promoter of GaGL2 complex, corresponding to sharply reduced transcription of GaGL2. Our results support that cotton homologs to the genetic molecules regulating Arabidopsis trichome differentiation interacted in the epidermis of ovules and the redundant GaMYB23 serves as a negative regulator in fuzz-fiber patterning.

[Genetic analysis of fuzzless in cotton germplasm].

The present study was conducted to evaluate genetic analysis of fuzzless seed trait in cotton. One hundred and two upland cotton (G. hirsutum) and eighty-five island cotton (G. barbadense) were used to cross with the same lines, TM-1 (G. hirsutum) and Xinhai 13 (G. barbadense), respectively. Two different F1 populations obtained were assessed to specify the dominant and recessive inheritance of fiber fuzziness in these lines. Three F1 populations (Kuguangzi × TM-1, Luwuxu × TM-1, and SA65 × TM-1) displaying recessive fiber fuzziness inheritance were selected to construct the F2 population for a further genetic study of fuzzless seed trait. The results of this study indicated that (1) the same materials showed different quantities of fuzzy fiber in different environments. Less fuzzy fiber was found in Xinjian and Hainan compared to Anyang. Thus, the quantity of fuzzy cotton seed depends on ecological environment. (2) In upland cotton, the inheritance of fiber fuzziness was dominant for 26 accessions (25.49%), incompletely dominant for 8 accessions (7.84%), and recessive for 22 accessions (21.57%). The inheritance of fiber fuzziness in island cotton was dominant for 5 accessions (5.88%), incompletely dominant for 16 accessions (18.82%), and recessive for 9 accessions (10.59%). Analysis of F2 population indicated that the fiber fuzziness of Kuguangzi was controlled by two recessive complementary effect alleles. The fiber fuzziness of Luwuxu was controlled by two recessive additive effect alleles, and a single recessive gene controlled the same trait for SA65. Fiber fuzziness evaluation in cotton germplasm provides the genetic and basic information for cotton fiber development study and breeding.

[Degradation characteristics of transgenic cotton residues in soil by Fourier transform infrared spectroscopy].

After transgenic cotton residues incubated in soil 430 d, the contents and structural characteristics of soil humus fractions, fulvic acid, humic acid and humin were measured by potassium dichromate titrimetric method and Fourier transform infrared spectroscopy. The results showed that all soil humus fractions increased after the degradation of cotton residues, and the most relative increase was with humin and the least was with fulvic acid. Compared to their near-isogenic non-transgenic cottons, soil humus content for transgenic Bt cotton residue decreased, and that forr transgenic Bt+CpTI cotton Z41 was approximate, but that for transgenic Bt+CpTI cotton SGK321 increased. Infrared spectroscopy of fulvic acid, humic acid and humin showed the addition of cotton residue decreased the content of oxygenous groups, and increased the alkyl and amide groups. There were differences in the speed to form soil humus among three transgenic cottons. Transgenic Bt cotton was slower than its counterpart, transgenic Bt+CpTI cotton Z41 was approximate to its counterpart, but transgenic Bt+CpTI cotton SGK321 was faster than its counterpart.