Genetic control of somatic embryogenesis in cotton petiole callus cultures

Summary Three commercial varieties (Acala SJ-5, Coker 312 and Paymaster 303) and three exotic accessions (T1, T25 and T169) of cotton (Gossypium hirsutum L.) were tested for ability to undergo somatic embryogenesis. Sections of split petiole were cultured on 3 media and evaluated for embryogenesis after 180 days. Embryogenic T25 and Coker 312 plants were selected and crossed in a diallel with non-embryogenic Acala SJ-5, Paymaster 303, T1 and T169 plants. F₁, F₂ and BC₁ populations were generated and tested for embryogenesis on a medium of MS salts and vitamins (1962) plus (per liter) 4.0 mg NAA, 1.0 mg Kn, 30 g glucose, 100 mg myo-inositol, 2.0 g Gelrite and 0.75 g MgCl₂. Segregation for both occurrence and magnitude of embryogenesis was observed, suggesting the action of more than one gene.     

Production of fertile hybrid germplasm with diploid Australian Gossypium species for cotton improvement

The 17 wild Australian Gossypium species are distant diploid relatives of the commercial tetraploid cottons, G. barbadense L. and G. hirsutum L. They interest cotton breeders as a source of terpenoid-aldehyde-free seeds, a trait only found in five Australian Gossypium species. They elicit further interest because some species grow near current and projected cotton growing areas in Australia and thus could serve as unintentional recipients of transgenes from genetically engineered cotton cultivars. The utility of the wild Australian Gossypium species in cotton breeding depends on the ability to generate fertile hybrids, and to the extent this is possible under glasshouse conditions, it allows predictions regarding the probability that fertile hybrids between the transgenic cottons and spatially associated populations of wild species will arise without human manipulation. The Australian Gossypium species fall into three morphologically and cytologically distinct groups designated the C, G, and K genomes, The G-genome species hybridize most readily with G. arboretum (a diploid A-genome cultivated cotton), while the C- and K-genome species are more compatible with G. hirsutum (a tetraploid AD-genome cultivated cotton). These intergenomic hybrids are sterile, and the chromosome complement of the hybrids must be doubled prior to backcrossing to G. hirsutum. The only exceptions were four G. hirsutum × K-genome triploids, which exhibited limited female fertility when backcrossed to G. hirsutum. Two of the three diploid species geographically associated with commercial cotton fields (G. australe F. Mueller & G. rotundifolium Fryxell, Craven & Stewart) failed to produce hybrid progeny when pollinated with G. hirsutum pollen; the third species (G. sturtianum J.H. Willis) produced only 5 sterile triploids from 25 pollinations. Thus, the probability that wild species could serve as recipients of transgenes is functionally zero, especially in conjunction with the profound prezygotic barriers that separate the cultivated tetraploid cottons from their wild Australian relatives. Eighteen new fertile synthetic polyploids and 23 self-fertile derivatives of two synthetic hexaploids were produced. Synthetic tetraploids require greater effort to backcross than do synthetic hexaploids. These fertile hybrids represent a new avenue of introgression of genes from wild Australian Gossypium species into commercial cotton cultivars, an avenue limited only by the level of recombination. This revised version was published online in July 2006 with corrections to the Cover Date.     

Somatic hybrids between Gossypium hirsutum L. (4×) and G. davidsonii Kellog (2×) produced by protoplast fusion

Somatic hybrids were obtained from electrofused protoplasts derived from embryogenic suspension cultures of tetraploid cotton (G. hirsutum L. cv. Coker 201) and embryogenic callus of diploid wild cotton G. davidsonii. The regenerants were initially identified as hybrids by RAPD (random amplified polymorphic DNA) analysis. Subsequently, observation on chromosome counting, morphology and SSR (simple sequence repeat) confirmed the hybrid status. Cytological investigation of the metaphase root-tip cells of the regenerated plants revealed there were 74 to 84 chromosomes in the plants, close to the expected 78 chromosomes. SSR analysis revealed the regenerated plants contained specific genomic fragments from both fusion partners, further confirmed their hybridity. The morphology of the plants was intermediate between the two fusion partners. The regenerants were difficult to develop into mature plants because their roots browned and they wilted from the stem apex before forming 3 to 5 true leaves. The hybrid plants were transferred to soil by grafting in vitro onto rootstocks.     

A novel and efficient method for identifying cotton leafroll dwarf virus infection in upland cotton (Gossypium hirsutum)

We report the development of a novel and more efficient, rapid, cost-effective and simple technique than current PCR-based identification methods for screening cotton (Gossypium hirsutum) plants for the presence of cotton leafroll dwarf virus (CLRDV). This protocol takes advantage of the PACE (PCR Allele Competitive Extension) system and uses PCR amplification of cDNA, coupled with sequence-specific fluorescent probes to differentiate between infected and uninfected cotton plants. This procedure has the potential for application in detection of other RNA viruses in a variety of other crops, by using primers specific for the RNA-dependent RNA polymerase (RdRP) gene and a widely conserved housekeeping gene in the host organism; in this case, the G. hirsutum polyubiquitin gene (GhUB).     

Trend and Stability Analysis to Interpret Results of Long-Term Effects of Application of Fertilizers and Manure to Cotton Grown on Rainfed Vertisols

A field experiment was conducted from 1985–1986 to 2002–2003 on Vertisols under rainfed conditions to evaluate the effect of cropping systems and application of fertilizers and manure on seed cotton yield. To determine the long‐term effects, trends and stability analyses were performed. Soil samples (0–0.15 and 0.15–0.30 m) were collected at the end of year 18 and analysed for available P and AB‐DTPA extractable Zn. Among cropping systems, Asiatic diploid cotton (Gossypium arboreum) yielded 233 kg ha^?1 more seed cotton than the upland tetraploid cotton (Gossypium hirsutum). Yield trend was positive for G. arboreum compared with G. hirsutum. However, the slope was not significant. Stability analyses indicated overall higher yield stability for G. arboreum than G. hirsutum. Compared with monocropping G. hirsutum, G. hirsutum–sorghum (Sorghum bicolor L.) (H‐S) rotation was significantly more stable. Soil samples (0–0.30 m) of the manure‐amended plots had significantly greater P and Zn content (above the critical limit) compared to those receiving inorganic fertilizers alone. With regard to nutrient management practices, seed cotton yield was the highest for the integrated nutrient management (INM) treatment receiving a combination of organic and inorganic fertilizers. Among primary nutrients, the effect of P was significant while that of K was not. Balanced fertilizer application was significantly better than treatments receiving N and NK. Yield trends were, in general, not significant. However, a positive trend was noticed for treatments receiving manure compared to fertilizer alone. Stability analysis, on the other hand, indicated that the slopes were, in general, significant. Among the nutrients, mean yield response with and without P was 1007 and 672 kg ha^?1 respectively. Combined application of manure and fertilizer (INM) resulted in the highest mean yield response (1218 kg ha^?1) and the slope was highly significant (P < 0.004). In the manure‐amended plots, a better nutrient status probably imparted a greater degree of yield stability. The present study suggests that compared to trend analysis, stability analysis being sensitive as it recognizes the treatment × environment interaction, is a better option to interpret results from long‐term agronomic experiments.     

Comparative Study for the Relative Importance of Characters Contributing to Seed-Cotton Yield in American and Egyptian Cottons

The path coefficient analysis was employed to determine the contribution of boll number per plant, boll weight and seed index characters to the variation in seed-cotton yield per plant, in the American and Egyptian cotton varieties; McNair 220 and Giza 75 respectively. Data obtained from yield trials carried out at Fayoum Experimental Farm, Faculty of Agriculture, in two growing seasons, 1982 and 1983, were used in this analysis.
It was found that boll number per plant had the greatest contribution to seed cotton yield in both American and Egyptian cotton varieties. Selection for number of bolls/plant and boll weight in American Upland cotton accounts for about 75% of the yield. While in Egyptian cotton, number of bolls/plant and boll weight account for about 60% only of yield.
The arrangement of the components contributing to plant yield, according to their relative importance in McNair 220 variety was:
(1) The direct effect of bolls/plant. (2) The direct effect of boll weight. (3) The joint effect of number of bolls with boll weight. While in Giza 75 variety the arrangement was as follows:
(1) The direct effect of number of bolls per plant. (2) The joint effect of number of bolls with boll weight. (3) The direct effect of boll weight.     

Assessment of Gossypium sturtianum and G. australe as potential sources of Fusarium wilt resistance to cotton

When challenged with Fusarium oxysporum f. sp. vasinfectum (Fov) from vegetative compatibility groups (VCGs) 01111 and 01112 in glasshouse tests, Gossypium australe Mueller and Gossypium sturtianum Willis accessions showed a variety of disease responses ranging from highly resistant to highly susceptible. Under high disease pressure G. sturtianum accession Gos-5275 was significantly more resistant than the commercial G. hirsutum cultivars that are designated standards for Fusarium resistance by Australian cotton breeders. Under low disease pressure G. sturtianum accession Gos-5250 was more susceptible than a highly susceptible commercial cultivar. A series of glasshouse tests was performed at two locations (Indooroopilly, QLD. and Canberra, ACT), and under low and high disease pressure. In these tests, a hexaploid cross (Gos-5271) generated from a Fusarium-resistant G. sturtianum (Gos-5275) and a Fusarium-susceptible G. hirsutum L. (CPI-138969) was significantly more resistant to Fusarium wilt than its G. hirsutum parent. Thus G. sturtianum, with a diploid genome and a range of responses to Fov challenge, has the potential to provide the basis for the elucidation of the genetic basis of resistance to Fusarium wilt in cotton species. In addition, resistant accessions of G. sturtianum are identified as a potential source of Fusarium wilt resistance genes for cotton breeding. In the glasshouse tests used to assess the resistance of various Gossypium accessions to Fusarium wilt disease, the scoring of vascular browning was found to give a more reliable indication of disease severity than the scoring of foliar symptoms. This revised version was published online in August 2006 with corrections to the Cover Date.     

Characterization of resistance gene analogs from <Emphasis Type="Italic">Gossypium arboreum</Emphasis> and their evolutionary relationships with homologs from tetraploid cottons

Four cotton species (genus Gossypium) produce spinable fiber. The two diploid species of Asiatic origin, Gossypium arboreum and G. herbaceum, have been largely replaced by G. hirsutum. However, these diploid species are potentially a rich source of genes for the improvement of G. hrisutum, particularly in terms of providing resistance against biotic and abiotic stresses. As a first step towards understanding the mechanisms of resistance in cotton, we designed 24 non-degenerate primers based on resistance gene analogs (RGAs) cloned from G. hirsutum for screening a number of cotton species with the A and D genomes. Most of these RGAs are conserved on the A genome (G. arboreum), suggesting a bias towards this genome. The amplified RGAs from G. arboreum were cloned and their nucleotide and amino acid sequences compared with RGA sequences available in public databases. The majority of the RGAs identified were homologous to those isolated from G. hirsutum and G. barbadense, but their diversity was greater than expected at both the nucleotide and amino acid levels. These RGAs provide useful tools for the identification of full-length resistance genes from bacterial artificial chromosome and cDNA libraries.     

Effects of Genotypes and Plant Density on Yield, Yield Components and Photosynthesis in Bt Transgenic Cotton

Bt (Bacillus thuringiensis) transgenic cotton (Gossypium hirsutum L.), including the introduced, indigenous Chinese non‐hybrid and hybrid cotton, is spreading very rapidly in China. Agronomic and photosynthetic performance as well as the optimum plant density for planting Bt cotton may vary with genotypes. With three types of commercial Bt cotton varieties, two field experiments were conducted to study yield performance and leaf photosynthetic rate (Pn) during 2000 and 2001, and yield interaction between variety and plant density during 2001 and 2002 in Yellow River region in northern China. The first experiment showed that the indigenous Chinese Bt cotton significantly differed from the introduced Bt cotton (IBtC) in plant growth and yield components. As a result of manipulation of boll numbers, boll weight and lint percentage, there was no significant difference in lint yield between Chinese non‐hybrid Bt cotton (CBtC) and the IBtC, but two Chinese hybrid Bt cotton (HBtC) varieties exhibited significantly higher lint yield than all other varieties in either 2000 or 2001. Hybrid cotton SCRC15 showed a one‐peak curvilinear change in diurnal course of Pn throughout the growing season, while non‐hybrid cotton 33B and SCRC16 exhibited severe mid‐day depression in Pn in squaring, flowering or boll‐setting stage. The second experiment showed that the main effect of plant population on lint yield was not significant, and yield difference among all treatments was derived from varieties and the interaction between plant density and variety. The optimal plant densities in terms of lint yield for the introduced, indigenous CBtC and HBtC genotypes were 6.0, 4.5 and 3.0 plants m^?2 respectively.     

Relationship between molecular marker heterozygosity and hybrid performance in intra- and interspecific hybrids of cotton

The present study was conducted to investigate the relationship between parental molecular marker diversity and hybrid performance in both intra‐ and interspecific hybrids of cotton to evaluate the feasibility of predicting hybrid performance using molecular markers. Three cytoplasmic male sterile (CMS) lines were crossed with 10 restorer lines to produce 22 F₁ hybrids during 2003. Of 22 F₁s, 14 hybrids were intraspecific (Gossypium hirsutum × G. hirsutum) and eight interspecific (G. hirsutum × G. barbadense). These 22 F₁ hybrids and their parents were evaluated for yield and fibre quality traits at Zhejiang University, Hangzhou, China during 2004 and 2005. Genetic distances (GD) among the parents were calculated from 56 random‐amplified polymorphic DNAs (RAPD) and 66 simple sequence repeat (SSR) marker data, and their correlation with hybrid performance and heterosis were analysed. The parents could be discriminated into G. hirsutum and G. barbadense clusters by cluster analysis based on both RAPD and SSR markers data. The correlation (r = 0.503, P ≤ 0.05) was calculated between GD_rapd (GD based on RAPD markers) and GD_ssr (GD based on SSR markers). Correlation of GD with hybrid performance and heterosis differed considerably between intra‐ and interspecific hybrids. The correlation between GD and hybrid performance was non‐significant for most of traits within the hybrids of G. hirsutum species. However, it was significantly and positively correlated for fibre length, fibre strength and elongation in interspecific hybrids. The relationship between GD and heterosis was observed to be positively significant for boll weight within hybrids of G. hirsutum with significant and negative correlations for fibre length and elongation. In conclusion, the power of predicting hybrid performance using molecular markers in cotton is low. But, the relationship between SSR marker heterozygosity and hybrid performance can be used to predict fibre length during interspecific hybrid cotton breeding.     

Effects of chromosome 5sh from Gossypium barbadense L. on flower production in G. hirsutum L.

Cotton (Gossypium spp.) yield is directly determined by mature bolls that developed from squares and flowers. The first four to six weeks of flowering accounts for the majority of lint yield in upland cotton (G. hirsutum L.) for most cultivated areas of the southern USA cotton belt. In this study, we evaluated 13 cotton chromosome substitution lines (CS-B) and their chromosome specific-F₂ hybrids, TM-1, 3–79, and six cultivars for the number of flowers produced during the first four weeks of flowering. Results showed that CS-B05sh produced more flowers than TM-1 and 3–79 from 10 July to 5 August. The results suggest that when the short arm of chromosome 5 was substituted from 3–79 (G. barbadense L.) into TM-1 (G. hirsutum) a positive genetic association with flower numbers during this flowering period was exhibited. CS-B05sh had comparable flower numbers with three cultivars, Deltapine 90, Phytogen 355, and Stoneville 474 and more flowers than, Sure Grow 747, Sure Grow 125, and Deltapine 5415. Different patterns for additive and dominance effects on cumulative flowers were observed across weeks of flowering. Dominance effects were more apparent during the early part of the flowering period while additive effects were more apparent towards the end of the flowering period.     

Using molecular markers and field performance data to characterize the Pee Dee cotton germplasm resources

Knowledge of genetic relationships in crop breeding programs provides valuable information that can be used by plant breeders as a parental line selection tool. In Upland cotton (Gossypium hirsutum L.), the Pee Dee germplasm program represents one of the most historically significant Upland cotton breeding programs and is known as a key source of fiber quality genes for commercial cultivars. The foundation of the Pee Dee germplasm is known to represent an array of genetic diversity involving the hybridization of G. hirsutum L., G. barbadense L., and triple hybrid strains (G. arboreum L. × G. thurberi Todaro × G. hirsutum L.). In this study, we characterized genetic relationships within the Pee Dee germplasm collection using molecular marker and field performance data. Molecular marker and field performance data showed the Pee Dee germplasm collection still maintains useful amounts of genetic diversity. The methods described provide plant breeders of cotton and other crops a strategy to develop a parental line selection tool based on genotypic and phenotypic information. Cotton breeders can directly use the information provided to select specific Pee Dee germplasm parental line combinations based on genotypic (molecular marker) and phenotypic (field performance) information rather than relying on pedigree and phenotypic information alone.     

Molecular diversity of Egyptian cotton (<Emphasis Type="Italic">Gossypium barbadense</Emphasis> L.) and its relation to varietal development

Twenty-eight Egyptian cotton (Gossypium barbadense L.) genotypes (varieties and hybrids) were used for analysis of genetic diversity using DNA based markers (ISSR, SSR, and EST) and to study varietal development of cotton. The ISSR markers gave the highest percentage of polymorphic bands as well as polymorphic information content compared with the other molecular markers (i.e. EST and SSR markers). Using clustering analysis, no general clustering according to the pedigree history of the genotypes was observed. Using principal coordinate analysis (PCOORDA), cotton genotypes were separated by the first three principal coordinates (PC1, PC2, and PC3) accounting for 11.5, 8.6, and 7.2% of the total genetic variance, respectively. The cotton genotypes were distributed into three parts based on the first PC, each part containing a group of varieties having a common ancestor. ‘Giza 12’ variety was the common ancestor for the varieties included in the first part and ‘Ashmouni’ variety was the common ancestor for the varieties included in the second part, while both ‘Sakha 3’ and ‘Sakha 4’ varieties were common ancestors for the varieties included in the third part. The results of the PCOORDA also showed better resolution of the genetic diversity than cluster analysis especially in the illustration of the varietal development of cotton. That means that principal coordinate analysis can be strongly used either alone or in combination with cluster analysis to discuss both genetic diversity and varietal development in the cotton genotypes.     

中国棉花体细胞植株再生的基因型分析

我国开展棉花体细胞组织培养研究20多年来,已经成功的获得了戴维逊氏棉、雷蒙德氏棉、瑟伯氏棉、拟似棉、草棉、亚洲棉、海岛棉和陆地棉8个棉属的再生植株。其中陆地棉约占总数的70％,海岛棉约占20％。考查陆地棉栽培品种(系)的系谱亲缘关系,主要来源于斯字棉的材料约占26％、岱字棉约占23％、乌干达棉约占12％、珂字棉约占11％、爱字棉约占3％、福字棉约占1．5％。本文对我国再生植株品种(系)的差异进行了分析,并将开展棉花体细胞组织培养以来再生成功或研究利用的基因型加以整理,以期为进一步的研究提供参考。     

Interspecific hybridization in Gossypium L.: characterization of progenies with different ploidy‐confirmed multigenomic backgrounds

Cytological and molecular investigations were undertaken for parent and progeny derived from a trispecific line [2(Gossypium arboreum × G. anomalum) × G. hirsutum var. BWR], which was crossed with G. hirsutum var. JLH168. Cytomorphological analysis of the F₁ (G. arboreum × G. anomalum), its amphidiploid and progeny from trispecies hybrid showed distorted ploidy segregation with monovalents to hexavalents and high intergenomic (small A₂ and large B₁) allosynthetic chromosome pairing. Microsatellite analysis identified three fragments associated with G. arboreum and G. anomalum and six fragments associated with G. hirsutum in derivates of the trispecies line × G. hirsutum var. JLH168. Inter‐Retrotransposon Amplified Polymorphism (IRAP) analysis revealed fragments of G. arboreum and G. anomalum, only in F₁ and amphidiploid. Chromosomal association and microsatellite analysis of three progeny genotypes (i.e. haploid, hexaploid and tetraploid no. 1) confirmed that they share multigenomic background from the three cotton species (A₂, A_hD_h and B₁ genome). The interspecific hybrid cotton genotypes studied are likely to be useful for the introgression of genes from diploid species to commercial upland cultivars.     

Fine mapping of the <Emphasis Type="Italic">fg</Emphasis> gene using the interspecific cross of <Emphasis Type="Italic">Gossypium hirsutum</Emphasis> × <Emphasis Type="Italic">Gossypium barbadense</Emphasis>

Frego (fg) bract is an important agronomic trait in tetraploid cotton, which has been widely introduced into several cotton varities or lines in the past several years. In order to help us further understand the underlying molecular mechanism of frego bract development, a map-base cloning strategy was used to localize the fg locus. An F₂ population which comprised of 290 fg individuals derived from a cross of the multiple-marker line T582 (G. hirsutum, carrying the fg gene) with Hai7124 (G. barbadense) was constructed. Genetic linkage analysis was carried out to map of the fg locus with SSR and EST-SSR markers in tetraploid cotton. Genetic linkage analysis showed that the fg locus was flanked by the marker NAU3016 and NAU3172 on the long arm of chromosome 3, with the genetic distance of 0.3 cM and 4.7 cM, respectively. The information of fg locus provided the basic information for the final isolation of this important gene in tetraploid cotton, these marker information could be used in marker-assisted selection in cotton.     

Transformation of α-EXPA1 gene leads to an improved fibre quality in Gossypium hirsutum

The Expansin protein is known for its multifaceted roles in plant growth, especially cell walls. However, very few studies have been done so far to assess the effects of expansin genes on Cotton fibre development. The present study is a successful effort to fill this gap, where the α-EXPA1 gene transformed into a local cotton variety, Gossypium hirsutum, through Agrobacterium-mediated transformation under Gossypium hirsutum Seed coat and Fibre-specific promoter (GhSCFP). The transgenic cotton plants underwent molecular characterization and fibre trait evaluation. Our results indicated that α-EXPA1 showed an up-regulated expression during the transition phase of secondary cell wall synthesis and resulted in improving the fibre parameters, especially micronaire value. Transgenic cotton fibre also showed a finer twisting under the Scanning electron microscope (SEM) as compared to non-transgenic cotton fibre samples. The fibre production is influenced by more than nine thousand genes, and the fibre improvement cannot be just achieved through a single gene transformation. However, α-EXPA1 is one of the potential candidates for cotton fibre research as it significantly improved the cotton fibre.     

Genetic detection of node of first fruiting branch in crosses of a cultivar with two exotic accessions of upland cotton

Flowering time has biological and agricultural significance for crops. In Upland cotton (Gossypium hirsutum L.), photoperiodic sensitivity is a major obstacle in the utilization of primitive accessions in breeding programs. Quantitative trait loci (QTLs) analysis was conducted in two F₂ populations from the crosses between a day-neutral cultivar Deltapine 61 (DPL61) and two photoperiod sensitive G. hirsutum accessions (T1107 and T1354). Node of first fruiting branch (NFB) was used to measure relative time of flowering. Different flowering time genetic patterns were observed in the two populations. Two QTLs were found across five scoring dates, accounting 28.5 (qNFB-c21-1) and 15.9% (qNFB-c25-1) of the phenotypic variation at the last scoring date in Pop. 1107 (DPL61 by T1107); whereas, one major QTL (qNFB-c25-1) can be detected across five scoring dates, explained 63.5% of the phenotypic variation at the last scoring date in Pop. 1354 (DPL61 by T1354). QTLs with minor effects appeared at various scoring date(s), indicating their roles in regulating flowering at a lower or higher node number. Genetic segregation analysis and QTL mapping results provide further information on the mechanisms of cotton photoperiodic sensitivity. Part of a Ph.D. dissertation by senior author submitted to the Department of Plant and Soil Sciences, Mississippi State University, December 2007. Contribution of USDA-ARS in cooperation with the Mississippi Agric. and Forestry Exp. Stn. Journal paper J. 11276 of Mississippi Agric. and Forestry Exp. Stn.     

基于主成分分析的不同棉花品种低温萌发关键期研究

为明确低温为害棉花种子萌发过程的关键期,以新陆中46号、新陆早57号、新陆早74号和新陆早82号为试验材料,25℃为对照温度,通过室内发芽试验,研究12℃低温处理下4个棉花品种在种子萌发过程中种子活力指标变化和胚根生长情况。结果表明,12℃低温处理能降低棉花种子活力和抑制胚根生长,不同品种间耐低温能力存在差异,新陆早57号低温耐受力最强。7个测定指标间均存在正相关关系,发芽指数与活力指数、胚根鲜重与胚根干重间相关性最大,发芽势与胚根长、发芽指数与胚根长间相关性最小。不同棉花品种各处理主成分综合得分排序为T 2相似文献