陆地棉遗传图谱构建与纤维品质性状QTL定位

以陆地棉(Gossypium hirsutum L.)品种Bar19/1和Acala1517-77杂交的108个F2单株为材料,应用85个标记(70个SSR标记和15个AFLP标记)构建了总长为814cM的遗传图谱,覆盖棉花基因组的18.3%。该图谱包含25个连锁群,分别对应到17条染色体和4个未知连锁群。应用复合区间作图法分析了该组合的108个F2单株和F3家系纤维品质性状,从遗传图谱上检测到19个纤维品质数量性状基因座(QTL),包括5个纤维长度、6个纤维比强度、4个伸长率及4个马克隆值QTL,分别解释各性状表型变异的15.11%~28.45%、8.46%~24.51%、11.08%~27.55%和9.23%~42.21%。纤维长度和伸长率的QTL以部分显性为主,少数具有超显性,比强度QTL以加性和部分显性为主,4个马克隆值QTL中有3个表现为超显性。研究结果表明,陆地棉Bar19/1和Acala1517-77间多态性位点丰富,有利于构建高密度遗传图谱,纤维品质性状的QTL分析从分子水平上揭示了纤维品质的遗传基础。     

Occurrence and pathogenicity of Enterobacter sp. causing sprout decay and seedling stunting of upland cotton (Gossypium hirsutum L.)

A new sprout decay and seedling stunting disease of unknown aetiology in upland cotton (Gossypium hirsutum L.) affecting nearly 5%–10% of young seedlings was noticed in vertisols of central Vidarbha (Maharashtra state, India) in July of 2017. The bacterium was consistently isolated from diseased seedlings and identified with a polyphasic method of characterization, including morphological, physiological, biochemical and 16S rRNA gene sequence analysis. The bacterium strain CICR-MGMG1 was isolated from diseased plants identified as Enterobacter sp. Inoculation of healthy cotton seed with an axenic culture of strain CICR-MGMG1 isolated from diseased young seedling reproduced disease symptoms of yellowing, stunting and deformed growth similar to the symptoms reported from infected field condition. The strain CICR-MGMG1 was consistently isolated from both diseased seedlings and stunted plants. Thus, the pathogenicity test of Koch's postulates was confirmed with the bacterium Enterobacter sp. strain CICR-MGMG1 as the causal organism of sprout decay and seedling stunting. To the best of our knowledge, this is the first record of Enterobacter sp. causing sprouts decay and seedling stunting of cotton.     

Genomic analyses reveal the genetic basis of early maturity and identification of loci and candidate genes in upland cotton (Gossypium hirsutum L.)

Although upland cotton (Gossypium hirsutism L.) originated in the tropics, this early maturity cotton can be planted as far north as 46°N in China due to the accumulation of numerous phenotypic and physiological adaptations during domestication. However, how the genome of early maturity cotton has been altered by strong human selection remains largely unknown. Herein, we report a cotton genome variation map generated by the resequencing of 436 cotton accessions. Whole‐genome scans for sweep regions identified 357 putative selection sweeps covering 4.94% (112 Mb) of the upland cotton genome, including 5184 genes. These genes were functionally related to flowering time control, hormone catabolism, ageing and defence response adaptations to environmental changes. A genome‐wide association study (GWAS) for seven early maturity traits identified 307 significant loci, 22.48% (69) of which overlapped with putative selection sweeps that occurred during the artificial selection of early maturity cotton. Several previously undescribed candidate genes associated with early maturity were identified by GWAS. This study provides insights into the genetic basis of early maturity in upland cotton as well as breeding resources for cotton improvement.     

Indirect somatic embryogenesis and plant recovery from cotton (Gossypium hirsutum L.)

Summary We describe a tissue culture procedure for somatic embryogenesis and plantlet regeneration in cotton (Gossypium hirsutum L. cv. Coker 312). Callused explants or individual globular embryos were transferred to basal media to induce somatic embryogenesis. To determine characteristic early indicators of successful germination and conversion, we identified six types of embryos that developed on basal media. Two of the six embryo types, designated as tulip-shaped and trumpet-shaped, could undergo conversion in preliminary tests, whereas the others had little or no developmental potential. Several media treatments designed to enhance the maturation of globular somatic embryos failed to increase the fraction of embryos which matured to form recoverable types. In efforts to improve plantlet recovery, tulip-shaped embryos were used in limited trials to contrast the effects of chemical and physical desiccation treatments on germination and conversion. The selective use of tulip-shaped somatic embryos, coupled with partial desiccation, seems to have augmented plant recovery. Growth habit, flowering, seed set, and lint production of most of the regenerated plants were comparable to seed-derived plants grown under the same conditions. Partial research support was provided by state and federal funds appropriated to the Ohio Agricultural Research and Development Center, The Ohio State University.     

Effect of host genotype on indigenous bacterial endophytes of cotton (Gossypium hirsutum L.)

The purpose of this study was to determine if populations of indigenous bacterial endophytes in seed, stem and root tissue of cotton seedlings are influenced by host genotype. Growth chamber and field experiments were conducted to test the hypothesis that host genotype has an effect on endophytic bacterial populations in seed tissues and the developing cotton seedling. Initially, population densities of bacteria within seed of nine cotton cultivars were very low (i.e., 10 2.0 colony forming units seed^–1). However, after 4 days growth on water agar, population densities within developing radicles increased significantly (log₁₀ 2–5 colony forming units) and significant cultivar differences were found. Significant cultivar differences occurred for populations of endophytic bacteria and the composition of bacterial functional groups differed among cultivars in field-grown seedlings at 5, 8, and 15 days after planting. Differences in the ranking of cultivars occurred for endophytic populations recovered from seed and aseptically and field-grown radicle and seedling tissues. These results suggest that whether originating from seed or from soil, cotton plants are capable of immediately establishing a carrying capacity for communities of endophytic bacteria following seed germination. During germination and development of the seedling, there are genetic and possible morphological/physiological effects that contribute to significant differences in colonization of bacterial endophytes among cotton cultivars.     

陆地棉主要产量相关性状的SSR标记关联分析

高产优质育种是我国棉花育种的主要目标。寻找与目标性状关联的分子标记,可克服常规育种的盲目性,提高分子标记辅助选择育种的准确性。本研究对118份陆地棉种质资源的衣分、单铃重、单株铃数及子指等4个产量相关性状进行2年2点的表型鉴定,并利用覆盖全基因组的、有多态性的214对SSR标记进行标记与性状的关联分析。结果表明:118份材料的4个产量相关性状表型变异丰富,平均变异系数的变幅在6.1%~19.1%之间,且在各环境中表现较为稳定;基因型分析表明,214对标记共检测到460个等位变异,基因多样性指数平均为0.5151,PIC值平均为0.4587,表明该批标记具有较多的等位变异数和较高的基因多样性;群体结构分析表明该批材料可分为4个亚群,且各类群中材料与地理来源无对应关系;关联分析结果显示,在显著条件下(-log10P1.3,P0.05),共有39个标记位点能够在2个及2个以上的环境中同时检测到,其中有4个标记位点同时与2个以上性状相关联,进一步比较发现,有7个位点与前人研究结果一致,其余32个位点为新发现的位点。研究结果可为陆地棉产量性状遗传改良的分子标记辅助选择提供理论依据。     

Effects of ultraviolet-B radiation on cotton (Gossypium hirsutum L.) morphology and anatomy

Cotton (Gossypium hirsutum L.) crop, cultivated between 40 degrees N and 40 degrees S, is currently experiencing 2-11 kJ m-2 d-1 of UV-B radiation. This is predicted to increase in the near future. An experiment was conducted to study the effect of enhanced UV-B radiation on vegetative and reproductive morphology and leaf anatomy of cotton in sunlit, controlled environment chambers. From emergence to harvest, cotton plants were exposed to 0, 8 or 16 kJ m-2 d-1 of UV-B in a square wave approach for 8 h from 0800 to 1600 h. Changes in plant height, internode and branch length, mainstem node number, leaf area, length and area of petals and bracts, and anther number per flower were recorded. Epidermal cell and stomatal density, stomatal index, leaf thickness, and epidermal, palisade and mesophyll tissue thickness were also measured. Initial chlorotic symptoms on leaves turned into necrotic patches on continued exposure to enhanced UV-B. Exposure to high UV-B reduced both vegetative and reproductive parameters and resulted in a smaller canopy indicating sensitivity of cotton to UV-B radiation. Enhanced UV-B radiation increased epicuticular wax content on adaxial leaf surfaces, and stomatal index on both adaxial and abaxial leaf surfaces. Leaf thickness was reduced following exposure to UV-B owing to a decrease in thickness of both the palisade and mesophyll tissue, while the epidermal thickness remained unchanged. The vegetative parameters studied were affected only by high levels of UV-B (16 kJ m-2 d-1), whereas the reproductive parameters were reduced at both ambient (8 kJ m-2 d-1) and high UV-B levels. The study shows that cotton plants are sensitive to UV-B at both the whole plant and anatomical level.     

Measuring gene flow in the cultivation of transgenic cotton (Gossypium hirsutum L.)

Transgenic Bt cotton NewCott 33B and transgenic tfd A cotton TFD were chosen to evaluate pollen dispersal frequency and distance of transgenic cotton (Gossypium hirsutum L.) in the Huanghe Valley Cotton-producing Zone, China. The objective was to evaluate the efficacy of biosafety procedures used to reduce pollen movement. A field test plot of transgenic cotton (6×6 m) was planted in the middle of a nontransgenic field measuring 210×210 m. The results indicated that the pollen of Bt cotton or tfd A cotton could be dispersed into the environment. Out-crossing was highest within the central test plot where progeny from nontransgenic plants, immediately adjacent to transgenic plants, had resistant plant progeny at frequencies up to 10.48%. Dispersal frequency decreased significantly and exponentially as dispersal distance increased. The flow frequency and distance of tfd A and Bt genes were similar, but the pollen-mediated gene flow of tfd A cotton was higher and further to the transgenic block than that of Bt cotton (χ² = 11.712, 1 degree of freedom, p<0.001). For the tfd A gene, out-crossing ranged from 10.13% at 1 m to 0.04% at 50 m from the transgenic plants. For the Bt gene, out-crossing ranged from 8.16% at 1 m to 0.08% at 20 m from the transgenic plants. These data were fit to a power curve model: y=10.1321x ^−1.4133 with a correlation coefficient of 0.999, and y=8.0031x ^−1.483 with a correlation coefficient of 0.998, respectively. In this experiment, the farthest distance of pollen dispersal from transgenic cotton was 50 m. These results indicate that a 60-m buffer zone would serve to limit dispersal of transgenic pollen from small-scale field tests.     

The effects of NaCl on antioxidant enzyme activities in callus tissue of salt-tolerant and salt-sensitive cotton cultivars (Gossypium hirsutum L.)

Summary To determine NaCl effects on callus growth and antioxidant activity, callus of a salt-tolerant and a salt-sensitive cultivar of cotton was grown on media amended with 0, 75, and 150 mM NaCl. Callus of the salt-tolerant cultivar, Acala 1517-8 8, grown at 150 mM NaCl, showed significant increases in superoxide dismutase, catalase, ascorbate peroxidase, peroxidase and glutathione reductase activities compared to callus tissue grown at 0 mM NaCl. In contrast, callus tissue of the salt-sensitive cultivar, Deltapine 50, grown at 0, 75, and 150 mM NaCl, showed no difference in the activities of these enzymes. At the 150 mM NaCl treatment, peroxidase was the only antioxidant enzyme from Deltapine 50 with an activity as high as that observed in Acala 1517-88. The NaCl-induced increase in the activity of these enzymes in Acala 1517-88 indicates that callus tissue from the more salt-tolerant cultivar has a higher capacity for scavenging and dismutating superoxide, an increased ability to decompose H₂O₂, and a more active ascorbate-glutathione cycle when grown on media amended with NaCl.     

Changes in oxidative stress enzymes during artificial ageing in cotton (Gossypium hirsutum L.) seeds

The present study was carried out to elucidate the mechanism of seed deterioration in two cotton (Gossypium hirsutum L.) cultivars (HS6 and H1098). The seeds were artificially aged at 40 +/- 1 degree C and 100% relative humidity for 4 days. In both cultivars, germinability decreased, whereas membrane deterioration, as assayed by electrical conductivity of the seed leachates, increased progressively with artificial ageing. The decrease in germinability was well correlated with increased accumulation of total peroxide and malondialdehyde content and decreased activities of antioxidant enzymes peroxidase, catalase, ascorbate peroxidase, glutathione reductase and superoxide dismutase. Hydropriming for 2 h and ascorbic acid priming for 12 h partially maintained germination and the activities of various antioxidant enzymes under artificial ageing and the accumulation of peroxide and MDA content was decreased. The results suggest that cotton seed deterioration during accelerated ageing is closely related to a decrease in activities of various peroxide scavenging enzymes and to lipid peroxidation.     

Somatic embryogenesis and plant regeneration in cotton (Gossypium hirsutum L.)

Tissue culture methods for improvement of cotton has lagged seriously compared to other major crops. A method for regeneration of cotton which includes a morphogenetically competent cell suspension was needed to facilitate selection of stress-resistant variants and gene manipulation. Preliminary screening of eight strains of Gossypium hirsutum L. for embryogenic potential resulted in the production of somatic embryos in all strains. Coker 312 was selected for use in the development of a model regeneration system for G. hirsutum. Calli were initiated from hypocotyl tissues of 3-day-old-seedlings. Globular embryos were present after six weeks in culture. Calli were subcultured to liquid suspension in growth regulator-free medium. After three to four weeks, suspensions were sieved to collect globular and heart stage embryos. Collected embryos developed further when plated onto semi-solid medium. To induce germination and plantlet growth, mature embryos were placed on sterile vermiculite saturated with medium. Upon development of roots and two true leaves, plantlets were potted in peat and sand, and hardened. Mature plants and progeny have been obtained with this procedure. A high percentage of infertile plants was observed among the regenerants.Abbreviations NAA 1 naphthaleneacetic acid - IAA indole-3-acetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - MS Murashige and Skoog - BA 6 benzylamino purine - 2i P N⁶-(²-isopentenyladenine     

陆地棉产量性状QTLs的分子标记及定位

用我国的高产栽培品种泗棉3号和美国栽培品种TM-1为材料,构建F₂和F_2∶3作图群体,应用301对SSR引物和1040个RAPD引物,对产量性状QTLs进行了分子标记筛选,结果共筛选出了37对SSR多态性引物和10个RAPD多态性引物的49个位点,鉴定出了控制产量性状变异的主效QTLs。定位于第9染色体的连锁群,分别具有控制铃重、衣分和籽指的主效QTLs,铃重的2个QTLs分别解释F_2∶3群体表型变异的18.2%和21.0%;在F₂群体检测到的1个衣分QTL解释表型变异的25%,另一个衣分QTL在F₂群体和F_2∶3群体都检测到,解释F₂群体衣分的24.9%的表型变异,解释F_2∶3群体衣分的5.9%的表型变异;在F_2∶3群体铃重的一个QTL的同一位置同时检测到一个籽指QTL,它解释15.6%的表型变异,是一因多效或是紧密连锁的两个QTLs,有待进一步研究。本研究标记的产量性状主效QTLs可用于棉花产量性状的标记辅助选择。     

Current status of genetic engineering in cotton (Gossypium hirsutum L): an assessment

Abstract

Cotton is considered as the foremost commercially important fiber crop and is deemed as the backbone of the textile industry. The productivity of cotton crop, worldwide, is severely hampered by the occurrence of pests, weeds, pathogens apart from various environmental factors. Several beneficial agronomic traits, viz., early maturity, improved fiber quality, heat tolerance, etc. have been successfully incorporated into cotton varieties employing conventional hybridization and mutation breeding. Crop losses, due to biotic factors, are substantial and may be reduced through certain crop protection strategies. In recent years, pioneering success has been achieved through the adoption of modern biotechnological approaches. Genetically engineered cotton varieties, expressing Bacillus thuringiensis cry genes, proved to be highly successful in controlling the bollworm complex. Various other candidate genes responsible for resistance to insect pests and pathogens, tolerance to major abiotic stress factors such as temperature, drought and salinity, have been introduced into cotton via genetic engineering methods to enhance the agronomic performance of cotton cultivars. Furthermore, genes for improving the seed oil quality and fiber characteristics have been identified and introduced into cotton cultivars. This review provides a brief overview of the various advancements made in cotton through genetic engineering approaches.