转基因抗虫棉风险性分析及应对策略

主要针对转基因抗虫棉抗虫的时空性问题,棉花不同生育期、植株部位、不同害虫世代抗虫性的变化,抗虫棉对棉田害虫和天敌种群结构的影响,害虫对抗虫棉毒蛋白的抗性,抗虫棉抗性基因的漂移以及抗虫棉子产品的生物安全性等问题进行了分析,进而提出了降低转基因抗虫棉潜在生态风险性的措施。     

Field evaluation of insect-resistant transgenic Populus nigra trees

The performance of insect-resistant transgenic poplar trees (Populusnigra) expressing a Cry1Ac gene from Bacillus thuringiensis subsp. Kurstaki HD-1 against poplar defoliators was evaluated in the field at the Manas Forest Station in Xinjiang Uygur Autonomous Region during1994–1997. The results showed that the average percentage of highly damaged leaves on the transgenic trees was 10% while that on the control trees in nearby plantations reached 80–90%. The average number of pupae per m² of soil at 20cm depth in transgenic poplar plantation was 18 which was only 20% of that found in the non-transgenic control field. The number of pupae and the leaf-damage on transgenic trees described above are all far below the threshold set for chemical protection measures. The non-transformed poplar trees grown in the same plantation with the transgenic trees were also protected indicating that cross protection occurred between these two kinds of trees. Insect-resistant transgenic poplar trees have a potential application value in afforestation. This revised version was published online in August 2006 with corrections to the Cover Date.     

转基因抗虫棉Bt基因的遗传连锁分析

对中美两国所获得的转基因抗虫棉品种GK— 1 2、新棉 33B分别进行了 Bt基因的连锁测验。分析表明两个品种的试验结果相似 ,Bt基因未整合在棉花第 I、II、III、IV、V、VI、VII、IX、X、XI、XII、XIII、XVII连锁群及 cu、pg、gl1等基因所涉及的染色体上。阐明了 Bt基因染色体图谱定位的复杂性 ,并提出以原位杂交技术实现 Bt基因在棉花染色体组上的物理标图     

转基因抗虫棉栽培管理措施

目前生产上大面积种植的转Bt基因抗虫棉有着明显不同于常规棉的特征特性，如棉种发芽出苗顶土能力、苗期长势较弱、现蕾速度和蕾铃转化快、对钾肥和生长调节剂较敏感、后期易早衰等，因此转Bt基因抗虫棉有着不同于常规棉的配套栽培技术。     

Effects of fiber wax and cellulose content on colored cotton fiber quality

Forty-eight colored cotton lines and hybrids were studied on the wax, cellulose content and fiber quality. The result were as follow: The wax content of green cotton was five to eight times greater than that of white cotton. As for the brown cotton lines, the darker the brown fiber color, the greater the content of the fiber wax. The wax content of dark brown cotton was two times greater than that of white cotton. However, the content of the cellulose of white cotton was the greatest, next was that of the brown cotton, and the content of the cellulose of green fiber was the lowest. There were differences in fiber length, fiber strength, and fiber uniformity, not only among different varieties with the same color fiber, but also among different coloring types. The content of cellulose had a significantly negative correlation with the content of wax and fiber elongation, but significantly positive correlations between the content of cellulose and fiber length, fiber strength, fiber fineness, lint index, boll weight, and fiber uniformity were observed in the color cotton lines used in this experiment. The study on wax and cellulose contents will give constructive advice for the breeding and molecular research of the colored cotton, and help increasing the color stability of the fiber and its textiles.     

转Bt基因抗虫棉对非靶标昆虫的影响

本文就转Bt基因抗虫棉对非靶标害虫、天敌和经济昆虫的影响以及影响因素进行了论述。研究表明,转Bt基因抗虫棉使得一些对Bt蛋白不敏感的次要植食性非靶标害虫种群数量明显增加或呈加重趋势,但没有证据表明转Bt基因棉对棉田其它害虫的发生有直接的促进作用。外源Bt基因导入棉花后,引起的棉花重要营养物质含量的变化以及棉花叶片形态结构的改变可能会直接影响棉花害虫种群的数量,且转Bt基因棉田更有利于其它植食性昆虫的发生和危害。害虫种群数量和Bt蛋白也会直接或间接影响天敌的数量。目前大多数的研究表明,转Bt基因抗虫棉对家蚕和蜜蜂等经济昆虫无明显不利影响。     

Field evaluation of transgenic Brassica napus lines carrying a seed-specific chimeric 2S albumin gene

The agronomic performance of transgenic Brassica napus lines carrying chimeric 2S albumin seed protein genes was studied under field conditions. A total of 37 transgenic lines were compared to non-transformed lines. Plant development, morphological characteristics, yield parameters and seed composition were considered. Only a few transgenic lines differed from the control, and then only for particular parameters: four lines had altered leaf morphology, two differed in germination efficiency, one differed in yield, and three differed in some parameters of seed composition. These differences were not correlated with the presence or expression of the transgene and, thus, were more probably linked to cell- or tissue-culture effects, or to residual variation present in cv. ‘Drakkar’. A statistical interaction between variations in a limited number of seed composition parameters and cell haploid culture was observed. While the expression of the chimeric 2S albumin gene was not as high as hoped, this study demonstrates that transgenic lines can be isolated in which the intrinsic morphological and agronomic properties of the original lines are maintained.     

棉纤维品质改良的分子生物学基础

本文简要地评述了棉纤维特异性表达基因和棉花纤维素合成酶基因的克隆及其表达方面的研究进展。与棉纤维伸长有关的基因主要有GhCAP和pGhEXl,与棉纤维次生壁加厚有关的基因主要有H6、Fb-E6和FSl8A等,但这些基因的精确功能还有待进一步研究。棉花celA是从第一个高等植物中克隆出的编码纤维素合成酶催化亚基的基因。利用转基因技术将抗虫、抗除草剂基因和这些与纤维发育有关的基因导入棉花,不但可改良棉花的产量和纤维品质,降低生产成本,而且可减轻对环境的不利影响,从而增强棉花的市场竞争力。     

Quantitative analysis and QTL mapping for agronomic and fiber traits in an RI population of upland cotton

Genetic mapping is an essential tool for cotton (Gossypium hirsutum L.) molecular breeding and application of DNA markers for cotton improvement. In this present study, we evaluated an RI population including 188 RI lines developed from 94 F₂-derived families and their two parental lines, ‘HS 46’ and ‘MARCABUCAG8US-1-88’, at Mississippi State, MS, for two years. Fourteen agronomic and fiber traits were measured. One hundred forty one (141) polymorphic SSR markers were screened for this population and 125 markers were used to construct a linkage map. Twenty six linkage groups were constructed, covering 125 SSR loci and 965 cM of overall map distance. Twenty four linkage groups (115 SSR loci) were assigned to specific chromosomes. Quantitative genetic analysis showed that the genotypic effects accounted for more than 20% of the phenotypic variation for all traits except fiber perimeter (18%). Fifty six QTLs (LOD > 3.0) associated with 14 agronomic and fiber traits were located on 17 chromosomes. One QTL associated with fiber elongation was located on linkage group LGU01. Nine chromosomes in sub-A genome harbored 27 QTLs with 10 associated with agronomic traits and 17 with fiber traits. Eight chromosomes in D sub-genome harbored 29 QTLs with 13 associated with agronomic traits and 16 with fiber traits. Chromosomes 3, 5, 12, 13, 14, 16, 20, and 26 harbor important QTLs for both yield and fiber quality compared to other chromosomes. Since this RI population was developed from an intraspecific cross within upland cotton, these QTLs should be useful for marker assisted selection for improving breeding efficiency in cotton line development. Paper number J1116 of the Mississippi Agricultural and Forestry Experiment Station, Mississippi State University, Mississippi State, MS 39762. Mention of trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by USDA, ARS and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

转基因抗虫棉新品种华M2

华M2为湖北华之夏种子有限责任公司选育的转基因抗虫常规棉新品种,其早熟性较好,纤维品质优,耐枯萎病、黄萎病,丰产性好,于2020年通过湖北省农作物品种审定委员会审定,审定编号:鄂审棉20200003。     

Physiological and biochemical dissection of fiber development in colored cotton

Colored cotton is a very attractive proposition for the textile industry as it reduces the cost for dying. However, inferior fiber quality and non uniform color make it unsuitable for heavy machine spinning. Due to this reason cultivation of colored cotton on commercial scale has been restricted by cotton growers. The current study was planned to unveil the process of fiber development in isogenic lines of brown, green and white cotton by comparing their fiber length, flavonoid, cellulose, carbohydrate and enzyme activities (SuSy, SPS, acid invertase, Cox, NAD+, NADH, NADP+, and NADPH). White cotton fiber exhibited high cellulose contents (915.81 mg g^?1) and long fibers (3.07 cm) as compared to colored cotton fibers with low cellulose content (787.63 and 780.66 mg g^?1) and shorter fiber length (2.54 and 2.48 cm). Similarly the amount of flavonoids also varied significantly with maximum concentration (8.67 and 7.13 mg g^?1) in brown and green cotton fiber at 5 DPA as compared to white cotton fiber (3.12 mg g^?1) at 0 DPA. This high concentration of flavonoid not only lowered the total amount of carbohydrates but also lowered the sucrose transformation rate to developing cotton fiber in colored cotton. The activity of different enzymes (AI, SuSy and SPS, Cox, NAD+/NADH and coenzymes NADP+/NADPH involved in the metabolism of carbohydrates was higher in brown cotton fiber than green cotton fiber. However, high activity of these enzymes did not always correlated with higher amount of cellulose thus indicated the involvement of complex mechanisms for cellulose and flavonoid synthesis during the development of cotton fiber.     

基于GYT双标图对西北内陆棉区国审棉花品种的分类评价

西北内陆棉区是我国最重要的主产棉区,对该棉区历年国审棉花品种进行科学分类和综合评价有利于国审品种资源的合理利用和棉花生产效率的提升。本研究采用GYT双标图分析对2003—2019年期间西北内陆棉区37个国审棉花品种的产量与霜前花率、纤维长度、比强度、马克隆值、枯萎病指数和黄萎病指数等性状的组合水平进行了综合分析和品种分类评价。结果表明,西北内陆棉区37个国审棉花品种可划分为3个特征明显的品种类型。其中,I型品种包括新陆早13号、中棉所49、新陆早21号、巴13222、新46、天云0769、Z1112、新石K18、J206-5、新石K21、禾棉A9-9、创棉508、H33-1-4、金科20、新K28、创棉512和J8031等17个品种,是产量与其余性状组合协调最好的品种类型,在生产上推广应用价值最高。II型品种产量和纤维品质表现一般,抗病性差,在当前生产上应用价值有限。III型品种的抗枯萎病性表现最好,但在其余性状上表现略差,综合生产应用价值有限,也许可作为抗病亲本应用。同时,根据各品种的理想指数筛选出创棉512、J8031、新K28、H33-1-4、金科20、新陆早13号和创棉508等综合表现优良的品种,也鉴别出新陆棉1号、新陆早33号、创棉501号和新陆早51号等综合表现相对略差的品种。本研究采用的GYT双标图分析方法是基于“产量-性状”组合水平对品种进行综合评价和分类研究,产量-性状组合之间的相关关系更加简单,多数组合间表现显著正相关,更适用于品种的多性状直观选择和综合评价,通常比先前的GT双标图解释的变异比率高,双标图模型拟合度更好,结果更可靠。本研究采用GYT双标图分析方法对西北内陆棉区国审品种的分类研究揭示了该棉区国审棉花品种的分类特征和应用价值,为本区的国家棉花品种试验审定提供了借鉴,也为其他作物品种的类似研究提供了范例。     

AFLP marker associations with agronomic and fiber traits in cotton

DNA markers linked with major QTL contributing to traits of importance will be a useful tool for cotton (Gossypium spp.) genetics and breeding. We crossed four photoperiod-sensitive accessions of cotton, G. hirsutum L., with a cultivar, selected day-neutral plants and backcrossed four times to each of the four photoperiod-sensitive accessions, selecting day-neutral plants at each generation. The day-neutral plants from the first cross and the four backcross generations were advanced to the F₆. These 20 day-neutral lines and four cultivars were grown in two environments at Mississippi State, MS and scored for seven agronomic and fiber quality traits. They were also scored for AFLP markers using a bulk sample of leaves from each of 24 lines. More than 50 AFLP markers were associated with the seven traits with fewer markers associated with fiber than agronomic traits. However, one to four markers were associated with 22–93% of the phenotypic variability of each of the seven traits. The results suggest that selected markers could be used in marker assisted selection (MAS) in crosses designed to use alleles from exotic accessions or cultivars to develop elite breeding lines for cotton improvement.     

基因工程改良棉纤维研究进展

棉纤维是由胚珠表皮细胞经过分化、伸长、次生壁加厚等一系列过程发育形成的 ,其发育过程受基因表达控制。随着棉纤维发育分子生物学研究的深入 ,人们认识到通过遗传工程途径修饰棉纤维发育进程、改良纤维品质的可能性愈来愈大。现就近几年来基因工程改良棉纤维品质方面的研究进展作一简要介绍。     

Molecular mapping and characterization of traits controlling fiber quality in cotton

Cotton (Gossypium spp) is the world's leading natural fiber crop. Genetic manipulation continues to play a key role in the improvement of fiber quality properties. By use of DNA-based molecular markers and a polymorphic mapping population derived from an inter specific cross between TM-1 (G. hirsutum) and 3-79 (G. barbadense), thirteen quantitative trait loci (QTLs) controlling fiber quality properties were identified in 3-79, an extra long staple (ELS) cotton. Four QTLs influenced bundle fiber strength, three influenced fiber length, and six influenced fiber fineness. These QTLs were located on different chromosomes or linkage groups and collectively explained 30% to 60%of the total phenotypic variance for each fiber quality property in the F₂ population. The effects and modes of action for the individual QTLs were characterized with 3-79 alleles in TM-1 genetic background. The results indicated more recessive than dominant, with much less additive effect in the gene mode. Transgressive segregation was observed for fiber fineness that could be beneficial to improvement of this trait. Molecular markers linked to fiber quality QTLs would be most effective in marker-assisted selection (MAS) of these recessive alleles in cotton breeding programs. This revised version was published online in August 2006 with corrections to the Cover Date.     

Constitutive expression of Cry proteins in roots and border cells of transgenic cotton

Transgenic cotton plants expressing Cry1Ac and Cry2Ab, from the soil bacterium Bacillus thuringiensis (Bt), provide effective control of certain lepidopteran pests, however, little is known about the proteins below ground expression. We used ELISA to quantify in vitro expression of the Cry1Ac and Cry2Ab proteins in mucilage, root border cells and root tips in five transgenic cultivars of cotton compared to conventional cultivar Sicot 189. Expression of Cry proteins in roots and border cells of the transgenic cotton cultivars was constitutive and at detectable levels, with Cry1Ac and Cry2Ab protein expression ranging from <20 ppb to >100 pbb. To determine if genetically modified cotton demonstrated simple differences in properties of the root, when compared to an elite parental line (cv. Sicot 189), we enumerated border cells on seedling radicles. Border cell counts of 14 cultivars ranged from 0.2 to 1.1 × 10⁴ cells per root tip with an average of 5 × 10³ border cells. Border cell production in the transgenic cultivars was generally similar to that of both donor and elite parents, the exception being the cultivar Sicot 189, which had substantially more border cells than all of its transgenic derivatives. Comparison of border cell number with varietal disease resistance ranking found a limited relationship (r ² = 0.65, n = 7) between border cell numbers and the commercial resistance rank against Fusarium wilt of cotton. The implications of differences in cotton cultivar border cell number and root tip expression of Cry proteins for plant–microbe interactions in the rhizosphere and the soil ecosystem are yet to be resolved.     

Greenhouse evaluation and inheritance analysis of transgenic cotton expressing <Emphasis Type="Italic">PTM3</Emphasis>, a MADS-box gene of Aspen

The PTM3 (MADS-box) gene characterized from flower of Aspen was ectopically expressed in cotton and was found to result in desirable agronomic traits. Among several transgenic lines, the PTM3 cotton event-10 was found to flower significantly earlier than the control and yield better [Ramachandran et al. (2011)]. We report our findings on performance based on greenhouse evaluation and inheritance of PTM3 cotton event-10. The T₁ progeny of event-10 were grown in pots under RBD design. The progeny were confirmed by kanamycin imbibition test, PCR, and GUS assay which showed the segregation of transgene at about 3:1 ratio. GUS assay was performed with pollen from all transgenic plants; plants that expressed gus in all pollen versus those that showed segregation for expression were found to be at a ratio of 1:2. Similar to observations made in the T₀ generation of event-10, the agronomic evaluation of T₁ progeny exhibited, on an average, earliness of 13 days in flowering, 13.5 days in crop maturity, and 22% of yield enhancement. The T₂ progeny of homozygous lines were grown on field soil in the greenhouse under strip trial design to see the effect as observed from potted plants of T₁ progeny. Inheritance of transgene cassette to T₂ progeny was confirmed by the same tests used to analyze T₁ progeny. As exhibited by T₀ and T₁ progenies, the T₂ progeny also showed earliness of 8 days in flowering, 12 days in crop maturity, and 17% of yield enhancement. The data generated from the progeny over two generations confirms that the PTM3 cotton event-10 is superior in agronomic characters as compared to non-transgenic cotton and is of interest for breeding shorter duration varieties with improved yield.     

Targeted identification of association between cotton fiber quality traits and microsatellite markers

Primitive and exotic accessions of cotton are potential sources of favorable alleles for genetic improvement, enriching diversity in the genetically constricted gene pool of elite cultivars. Three exotic accessions of cotton (MDN101, MDN063 and MDN257), collected from different parts of Central America and converted to day-neutral flowering; and four elite cultivars (PD94042, DES56, PMHS200 and Acala Maxxa) representing the US cotton gene pool were used as parents to create experimental populations. The corresponding F₂ and F_2:3 progenies of these populations were grown in two successive years (i.e., some in 2011–2012, some in 2012–2013) and phenotypes were scored in both F₂ and F_2:3 progenies in all 3 years (2011–2012–2013). These populations were screened with 113 polymorphic microsatellite markers selected from “hotspots” for fiber quality quantitative trait loci in the cotton genome and single marker analyses were performed to identify significant associations of the markers with six fiber quality traits. A total of 134 nominal marker-trait associations were identified, among which 15 were significant after Bonferroni correction for multiple comparisons. In 67 of 134 nominal associations and 4 of 15 significant associations, the exotic parents contributed favorable alleles to multiple backgrounds and for multiple traits, in addition to the traits for which they were selected. These results indicate that utilization of exotic and wild accessions of cotton is useful in introducing favorable alleles into the cultivated cotton gene pool for genetic improvement.