Gene effects for fibre properties in upland cotton (Gossypium hirsutum L.)

Summary Six populations — P₁,P₂,F₁,F₂,B₁ and B₂ — each of five Upland Cotton (Gossypium hirsutum L.) crosses were used to evaluate gene effects in the inheritance of fibre properties by Gamble's six-parameter model for the analysis of generation means. Partial dominance of long fibres over short fibres and of mature fibres over immature fibres was found in the material studied. Overdominance in gene action governed fibre fineness while additive gene action governed the fibre strength. Besides additive and dominance effects, significant epistasis was present in all crosses. These results indicate a significant potential for improving fibre properties through reciprocal recurrent selection.     

Inheritance of the fuzzless-lintless character in cotton (Gossypium hirsutum)

Summary A fuzzless-lintless mutant was identified in MCU.5 (Gossypium hirsutum) cotton in 1984. The inheritance of this character is reported in this paper. The fuzzless-lintless mutant was crossed with fuzzy-linted parents viz. MCU.5, MCU.7, Express Sindh (W), Piedmont Cleveland and Sindis Wild and the segregation pattern was studied in F₂ and BC₁F₁ generations. The segregation ratios for fuzzy-linted and fuzzless-lintless were 151 in the first cross, 631 in the second, third and fourth crosses and 2551 in the fifth cross. These ratios indicated that this character is controlled by 2–4 gene pairs, and the fuzzless-lintless character is a recessive to fuzzy-linted character. The chi-square test was significant only in the BC₁F₁ generation with MCU.7 and Express Sindh (W). The test revealed that the observed values deviated significantly from the expected ratio of 71, suggesting that this character is also influenced by modifier gene complex.     

Correlation studies on yield and fibre traits in upland cotton (Gossypium hirsutum L.)

Summary Two diverse parents of upland cotton namely J.34 and I.C. 1926 were crossed. A comparison between biparental intermated progenies and F₃ families indicated alteration of correlation coefficient between yield and halo length. The significant negative correlation in F₃ population between these two attributes changed to a positive but non significant one in biparental intermated progenies. A change in correlation coefficients was expected due to breakage of linkage upon intermating. An increase in the correlation coefficients could also be expected when linkages are predominantly in the repulsion phase. It is suggested that intermating in early generations coupled with selection of desirable segregants may prove a useful method for improving yield and quality simultaneously. The diallel selective mating system may also supplement intermating to improve yield and quality in cotton.Part of Ph.D. Thesis submitted to the Haryana Agricultural University. Hissar-125004, India     

Isolation and characterization of terpene synthases in cotton (Gossypium hirsutum)

Cotton plants accumulate gossypol and related sesquiterpene aldehydes, which function as phytoalexins against pathogens and feeding deterrents to herbivorous insects. However, to date little is known about the biosynthesis of volatile terpenes in this crop. Herein is reported that 5 monoterpenes and 11 sesquiterpenes from extracts of a glanded cotton cultivar, Gossypium hirsutum cv. CCRI12, were detected by gas chromatography–mass spectrometry (GC–MS). By EST data mining combined with Rapid Amplification of cDNA Ends (RACE), full-length cDNAs of three terpene synthases (TPSs), GhTPS1, GhTPS2 and GhTPS3 were isolated. By in vitro assays of the recombinant proteins, it was found that GhTPS1 and GhTPS2 are sesquiterpene synthases: the former converted farnesyl pyrophosphate (FPP) into β-caryophyllene and α-humulene in a ratio of 2:1, whereas the latter produced several sesquiterpenes with guaia-1(10),11-diene as the major product. By contrast, GhTPS3 is a monoterpene synthase, which produced α-pinene, β-pinene, β-phellandrene and trace amounts of other monoterpenes from geranyl pyrophosphate (GPP). The TPS activities were also supported by Virus Induced Gene Silencing (VIGS) in the cotton plant. GhTPS1 and GhTPS3 were highly expressed in the cotton plant overall, whereas GhTPS2 was expressed only in leaves. When stimulated by mechanical wounding, Verticillium dahliae (Vde) elicitor or methyl jasmonate (MeJA), production of terpenes and expression of the corresponding synthase genes were induced. These data demonstrate that the three genes account for the biosynthesis of volatile terpenes of cotton, at least of this Upland cotton.     

Characterization of two cotton (Gossypium hirsutum L) invertase genes

Two cotton vacuolar-invertase genes were identified and sequenced. Both genes had seven exons, including an unusually small second exon typical of acid invertases. These genes encode peptides with many features shared by acid invertases from other species including, leader sequences that probably target the peptide to the vacuole, active site motifs and substrate binding motifs. Expression analyses indicated that one of the genes was expressed in roots during the starch filling stage of development. However, expression of the same gene fluctuated during the starch utilization stage of development. Therefore this gene was unlikely to play a role in determining sink strength of this tissue. Both genes were expressed in elongating fibers where they were likely to play a role in cell expansion. The invertase gene uniquely expressed in fiber had a simple sequence repeat (SSR) in the third intron that was polymorphic among various cotton species. An EST was identified with an expansion of the SSR that included the third intron indicating this SSR is associated with a splice variant. The polymorphic SSR may be useful in investigating the function of this gene in fiber development.     

Molecular insight into cotton leaf curl geminivirus disease resistance in cultivated cotton (Gossypium hirsutum)

Cultivated cotton (Gossypium hirsutum) is the most important fibre crop in the world. Cotton leaf curl disease (CLCuD) is the major limiting factor and a threat to textile industry in India and Pakistan. All the local cotton cultivars exhibit moderate to no resistance against CLCuD. In this study, we evaluated an exotic cotton accession Mac7 as a resistance source to CLCuD by challenging it with viruliferous whiteflies and performing qPCR to evaluate the presence/absence and relative titre of CLCuD‐associated geminiviruses/betasatellites. The results indicated that replication of pathogenicity determinant betasatellite is significantly attenuated in Mac7 and probably responsible for resistance phenotype. Afterwards, to decipher the genetic basis of CLCuD resistance in Mac7, we performed RNA sequencing on CLCuD‐infested Mac7 and validated RNA‐Seq data with qPCR on 24 independent genes. We performed co‐expression network and pathway analysis for regulation of geminivirus/betasatellite‐interacting genes. We identified nine novel modules with 52 hubs of highly connected genes in network topology within the co‐expression network. Analysis of these hubs indicated the differential regulation of auxin stimulus and cellular localization pathways in response to CLCuD. We also analysed the differential regulation of geminivirus/betasatellite‐interacting genes in Mac7. We further performed the functional validation of selected candidate genes via virus‐induced gene silencing (VIGS). Finally, we evaluated the genomic context of resistance responsive genes and found that these genes are not specific to A or D sub‐genomes of G. hirsutum. These results have important implications in understanding CLCuD resistance mechanism and developing a durable resistance in cultivated cotton.     

Breeding for reduced seed coat fuzz in Upland cotton (Gossypium hirsutum)

The inheritance of seed coat fuzz was studied in two half diallel sets of crosses of Upland cotton. One with F₄ selections from an inter-varietal cross showed a significant level of non-additive variance attributable to dominance and non-allelic interaction. In the other, using inbred varieties of diverse origin, the genetic control of seed fuzz was adequately accounted for by an additive-dominance model with no interaction. Genotypic correlations between seed coat fuzz, yield and lint quality characters, calculated for both diallel sets and for two other groups of breeding material, showed good agreement within each experiment between parents and hybrids or between parents and progenies but no consistent pattern between experiments. The results serve to emphasize the risks in extrapolating correlations from one group of breeding material to another. A useful level of reduced fuzz has been obtained in selections from the AH breeding programme and the genetical investigations indicate that a further reduction may be possible, thereby leading to easier handling of seed, speedier and cheaper ginning, low levels of seed coat nep and better seed germination.     

Polysaccharides in the culture medium of cotton (Gossypium hirsutum L.) ovules cultured in vitro

Polysaccharides secreted into the culture medium (PCM) from cotton (Gossypium hirsutum L.) ovules, culturedin vitro, have been examined. The amount of the polysaccharides increases with the duration of the culture, but except for the galactose content, does not vary markedly with the culture age. Analysis shows that the polysaccharide mixture contains mainly pectins, but there is also a significant amount of xyloglucan. The source of the polysaccharides is most likely the callus tissue which develops on the ovules.Abbreviations PCM polysaccharides secreted into the culture medium Presented at the Fourth Cell Wall Meeting, Paris, France (1986)     

A genetic approach to in vitro regeneration of non-regenerating cotton (Gossypium hirsutum L.) cultivars

Selections were made among individual plants of Gossypium hirsutum cv `Coker 310' for high-frequency in vitro regeneration by somatic embryogenesis. After three generations of selection, a pure line for high-frequency somatic embryogenesis was selected and named Coker 310 FR (FR, fully regenerating). Coker 310 FR could be regenerated by following previously published protocols (see Materials and methods) and a modified protocol developed in this study that reduced the time necessary for in vitro regeneration. Coker 310 FR was crossed with individual plants of major cotton cultivars grown in India, namely `MCU 5', `MCU 7', `Khandwa 2', `Bikaneri Nerma', `F 846' that have been shown to be recalcitrant to in vitro regeneration, to evaluate the regeneration potential of F₁s. All the F₁s showed regeneration by somatic embryogenesis. However, the F₁ of G. barbadense×G. hirsutum Coker 310 FR did not regenerate. Received: 16 September 1997 / Revision received: 1 April 1998 / Accepted: 15 May 1998     

Bemisia tabaci （Hemiptera： Aleyrodidae） nymphal feeding in cotton （Gossypium hirsutum） leaves

We used brightfield electron microscopy （BEM）, differential interference contrast microscopy （DICM）, scanning electron microscopy （SEM）, transmission electron microscopy （TEM）, and confocal laser scanning microscopy （CLSM） to investigate the stylet pathways of Bemisia tabaci during nymphal feeding behavior in cotton leaves beginning with penetration of the abaxial leaf surface and ending with stylets in sieve tubes in phloem tissues. Most nymphal stylets within salivary sheaths penetrating leaf tissues made complex turns and developed more than one salivary sheath branch before ending in sieve tubes. The external morphology of the salivary sheaths and their routes between and through leaf cells are described during the present study. Results showed the presence of the stylet within the sieve tubes. B. tabaci nymphs may remove stylets and feed in different sieve tubes. Ten short movies showing the progression of the stylet penetrations from adaxial surface to the sieve tubes are attached to Figures 8-15. The report and movies can be viewed from the internet. Download the movies to a local drive in your computer first for fast upload. The movies are posted on the website http：//www.ars.usda.gov/Services/docs.htm？docid=14629. The movies can be used as a teaching aid in biology classes.     

在自然衰老和诱导条件下棉花悬浮细胞程序性死亡的发生

Cotton suspension cells grew well in the MS medium supplemented with 0.1 mg/L 2,4 D and 0.1 mg/L KT. Senescence occurred when the cells were unsubcultured. The cells began to lose their viabilities on the 17th day, and on the 21th day oligonucleosomal sized DNA fragments ( DNA ladder) could be detected. Oligonucleosomal sized DNA fragments ( DNA ladder) was the hallmark of the programmed cell death. Programmed cell death of cotton suspension cells could be induced respectively by some stress factors which included heatshock (42+/-3 degrees C for 8 hours), 10 micromol/L camptothecin, 20 micromol/L fumonisin B1 and 50 mmol/L cycloheximide. The cotton suspension cells which grew in the MS medium supplemented with 0.1 mg/L 2,4 D and 0.1 mg/L KT differred physiologically from the cells in the MS medium supplemented with 0.1 mg/L IBA and 0.1 mg/L KT, and they responded differentially to the heatshock, 10 micromol/L camptothecin and 20 micromol/L fumonisin B1, while the same to 50 mmol/L cycloheximide.     

In vitro induction of multiple shoots and plant regeneration in cotton (Gossypium hirsutum L.)

Induction of multiple shoots in cotton (Gossypium hirsutum L. cv. Anjali-LRK 516) has been achieved with cotyledonary nodes devoid of cotyledons and apical meristems. Explants from 35-day-old seedlings yielded the maximum number of shoots (4.7 shoots/explant) using Murashige and Skoog (MS) basal medium supplemented with 6-benzylaminopurine and kinetin (2.5 mg/1 each). Explants from 35-day-old seedlings raised in glass bottles produced a higher number of multiple shoots (8.3 shoots/explant) than those grown in glass tubes and cultured on the same shoot induction medium. Elongation of multiple shoots was obtained on liquid or agar MS basal medium without phytohormones. In vitro shoots were rooted on half-strength agar-solidified MS basal medium or with 0.05 or 0.1 mg/1 naphthaleneacetic acid. Hardening and survival of tissue culture plantlets was 95% under greenhouse conditions.Abbreviations BAP 6-Benzylaminopurine - GA₃ Gibberellic acid - MS Murashige and Skoog medium - NAA -Napthaleneacetic acid     

Infraspecific DNA methylation polymorphism in cotton (Gossypium hirsutum L.)

Cytosine methylation is important in the epigenetic regulation of gene expression and development in plants and has been implicated in silencing duplicate genes after polyploid formation in several plant groups. Relatively little information exists, however, on levels and patterns of methylation polymorphism (MP) at homologous loci within species. Here we explored the levels and patterns of methylation-polymorphism diversity at CCGG sites within allotetraploid cotton, Gossypium hirsutum, using a methylation-sensitive amplified fragment length polymorphism screen and a selected set of 20 G. hirsutum accessions for which we have information on genetic polymorphism levels and relationships. Methylation and MP exist at high levels within G. hirsutum: of 150 HpaII/MspI sites surveyed, 48 were methylated at the inner cytosine (32%) and 32 of these were polymorphic (67%). Both these values are higher than comparable measures of genetic diversity using restriction fragment length polymorphisms. The high percentage of methylation-polymorphic sites and potential relationship to gene expression underscore the potential significance of MP within and among populations. We speculate that biased correlation of methylation-polymorphic sites and genes in cotton may be a consequence of polyploidy and the attendant doubling of all genes.     

Okra-leaf accessions of the upland cotton (Gossypium hirsutum L.): genetic variability in agronomic and fibre traits

Okra-leaf types of the upland cotton have the potential to be competitive to the normal-leaf types in yield and fibre quality, in addition to its potential resistance to insect pests and drought. Okra-leaf cotton accessions, collected at Cotton Research Institute, Faisalabad, Pakistan, were evaluated in respect of genetic variance and relative performance in half- and full-sib crosses (combining ability) for 2 years. Variation due to parents x years interaction was significant for lint percentage, seed weight and earliness index, resulting in moderately low but significant genetic variance across environments (years) for these traits. Interaction of environment with general combining ability was significant for seed yield, seed weight, and earliness index. General combining ability variation, contributed by females and males together, accounted for 71% of the total variation available for seed cotton yield, 60% of that for seed weight and height to node ratio each, and 75% of that for earliness index. Specific combining ability variation accounted for 85% and 51% of the total variation available for lint percentage and staple length, respectively. The contribution of female parents to general combining ability variation was higher than that of male parents for seed cotton yield, seed weight, height to node ratio, and earliness index. Okra-leaf accessions HR-VO-MS and HR107-NH were predicted to produce progenies having high yield, HR109-RT high lint percentage, while HR100-Okra, Gambo-Okra and HR-VO-1 were predicted to impart early crop maturity to their progenies by reason of their good general combining ability for these traits. The results also provided evidence that genes controlling high yield in HR-VO-MS and HR107-NH were different from those controlling high yield in HR109-RT. The set of genes controlling the high earliness index in HR100-Okra and that in HR-VO-MS also appeared to differ in expression.