共查询到17条相似文献，搜索用时 734 毫秒
1.
基因枪介导的海岛棉(Gossypium barbadense L.)茎尖遗传转化体系的建立 总被引：1，自引：1，他引：0
以新疆海岛棉品种新海13号的茎尖为转化受体,采用基因枪介导法,研究DNA包裹浓度、微载体种类、前渗处理时间、轰击条件、筛选方法等对茎尖转化的影响,建立了基因枪法介导的海岛棉茎尖转化体系。研究结果表明,采用1.5μg·μL-1的质粒DNA包裹1.0μm金粉微载体,前渗处理12 h,轰击压力为7.584 MPa(1100 psi),轰击距离为6 cm,后渗处理16 h,恢复处理24 h,在120 mg·L-1卡那霉素浓度下直接筛选35 d。在添加4g·L-1活性炭粒的生根培养基中诱导抗性幼苗生根,获得抗性再生植株。通过PCR和RT-PCR检测,共得到5株转抗除草剂基因EPSPS的阳性植株。该研究体系的建立为海岛棉的遗传转化奠定了基础,为海岛棉的育种工作提供了材料。 相似文献
2.
3.
以棉花茎尖分生组织为受体进行基因枪轰击转化的研究 总被引：3，自引：1，他引：3
以棉花茎尖分生组织为受体进行基因枪遗传转化,在不受宿主基因型范围和操作时间的限制等方面优于农杆菌介导法和花粉管通道技术。影响棉花茎尖分生组织基因枪遗传转化率的因素较为复杂,轰击前,外植体制备需考虑茎尖分生组织的取材时间、下胚轴保留的长度、外源生长调节剂及活性炭的使用量,以保证其正常生长;轰击后,抗生素筛选的浓度、筛选培养基的转换时间和抗性苗的壮苗也会影响转化效率;此外,DNA和钨粉或金粉的质量、基因枪参数及操作等对转化效果都有一定的作用。除对影响茎尖分生组织基因枪转化率的因素分析以外,还对近年来在棉花茎尖分生组织遗传转化中的最新动态进行了综述,并对目前存在的问题加以讨论。 相似文献
4.
5.
以茎尖分生组织为受体用农杆菌介导法转化激发子基因pemG1棉花植株的获得 总被引：1，自引：1，他引：0
为了解决棉花常规遗传育种周期长,农杆菌介导的转化下胚轴再生难,转化率低,胚变异率高等难题。通过改善茎尖的遗传转化,利用农杆菌介导法转化棉花茎尖,试图缩短转化周期,获得转化苗。构建了稻瘟菌激发子基因pemG1的植物表达载体pCAMBIA2300-pemG1,分别以‘珂字312’、‘中棉24’棉花茎尖分生组织作为外植体,摸索无菌苗的最佳培养方案,确定最佳共培养温度、共培养时间及卡那霉素筛选浓度。经过共培养,茎尖在诱导芽培养基上诱导生芽,从不同浓度的BAP和NAA生长激素的组合中,最终确定0.1 mg/L的BAP和NAA最适合芽的诱导;经过两个月的继代培养,在含有GA3的茎增殖培养基上完成茎的增殖和延伸;切取幼芽并使其在含有0.1 mg/L的IBA生根培养基中生根;生根的抗性植株移栽到温室中成苗,获得了T0代转基因株系4株。进行了转pemG1阳性植株的初步验证。 相似文献
6.
7.
基于抗草甘膦基因的棉花茎尖农杆菌介导转化方法的研究 总被引：1，自引：0，他引：1
以中棉所49、珂字棉201和YZ-1为受体材料,通过对培养基草甘膦浓度、农杆菌侵染时间和浓度、共培养时间以及恢复培养条件等因素的优化,建立了基于抗草甘膦基因的棉花茎尖农杆菌转化技术体系,并将抗草甘膦基因(EPSPS-G6)导入3个受体材料,获得转基因抗草甘膦棉花植株.研究结果表明,适合于棉花茎尖农杆菌介导的草甘膦筛选浓度为10 mg·L-1;茎尖转化体系为农杆菌菌液OD600为0.9～1.0,侵染时间为20min,共培养时间为48 h,选用SH培养基并加入适量活性炭(0.5 g·L-1)作为恢复培养基.用本研究创制的转化技术体系,转化处理3个陆地棉受体360个茎尖,共获得60株抗性再生植株,经PCR检测,获得阳性抗性植株26株,移栽成活23株.以茎尖外植体数计算,本体系的转化成功率6.4％.该转化体系适合于转化抗草甘膦基因或以抗草甘膦基因为筛选基因的外源基因,具有转化频率高、嵌合体少、转化周期短等优点. 相似文献
8.
9.
10.
11.
[Objective] The purpose of this study was to expand upland cotton genotypes suitable for regeneration via somatic embryogenesis, so as to provide excellent receptor materials for the genetic transformation of cotton. [Method] Six different hormone combinations were studied to establish a somatic embryo regeneration system for hypocotyl explants of Xinluzao 45, a widely planted variety in Xinjiang Province. [Result] Faint yellow callus was induced on a medium consisting of hormone combinations C1 (0.1 mg·L－1 acetic acid dichlorobenzene oxide [2, 4-D] + 0.1 mg·L－1 kinetin [KT]) or C5 (0.1 mg·L－1 2, 4-D + 0.5 mg·L－1 indo-3-butyric acid + 0.1 mg·L－1 KT). The generated callus was able to successfully induce embryogenic callus on hormone-free medium containing doubled KNO3. The embryogenic callus then developed into somatic embryos and regenerated plants. Maximum rates of callus induction and regeneration were 92.2% and 40.3%, respectively. Although no significant difference was observed in the rate of callus induction of Xinluzao 45 hypocotyls between hormone combinations C1 and C5, the incidence of induced embryonic callus was higher on C1 than on C5. [Conclusion] The somatic embryo regeneration system developed in this study provides technical support that will facilitate Agrobacterium-mediated transformation of upland cotton Xinluzao 45. 相似文献
12.
铜基营养保护剂对棉花生长和产量的影响 总被引：2，自引：0，他引：2
Lu Yanyan Zhang Min Tian Xiaofei Geng Jibiao Li Chengliang Ma Jinzhao Yang Jie 《棉花学报》2015,27(5):454-462
2013―2014年采用盆栽试验研究了不同种类及浓度的铜基营养保护剂对棉花生长、产量及防病效果等的影响。结果表明:4种铜基营养保护剂在喷施质量浓度为1 g·L-1和2 g·L-1时,与喷施清水相比,皮棉产量分别增加4.22%~13.38%和21.89%~38.36%;增加了棉株地上部干物质质量,最高达23.96%;降低了棉花病情指数26.75%~40.91%和22.78%~37.78%,但处理间差异不显著。因此,在棉花上喷施铜基营养保护剂既可促进棉花生长,增加棉花产量,又可提高棉花的防病效果。 相似文献
13.
We analyzed the effects of glyphosate to the index of salinization and alkalization by simulation in soils collected from cotton fields in Anyang, Henan and Anqing, Anhui, China, in 2014. The results showed that application of glyphosate changed electrical conductivity (EC), total alkalinity (TAL), exchangeable sodium percentage (ESP), sodium adsorption ratio (SAR) and pH of soil in both regions，compared with the blank control. With increased frequency of glyphosate application and increased concentration applied, all indices increased in the Anqing cotton fields soil which were treated with concentrations of 2.5-20 g·L－1 . However, EC, TAL and ESP increased dynamically in Anyang soil to which high concentrations of 20 g·L－1 were applied only four consecutive times. After different glyphosate treatments, the soil pH value showed an upward trend but smaller changes. Therefore, after repeated use of glyphosate to soils of two different region soils, although the changes of all indices were within the range of non-saline and non-sodic soils, the soils affected by salt in the glyphosate aqueous solution tended towards saline-sodic soils. 相似文献
14.
【目的】研究脱落酸(Abscisic acid, ABA)对棉花体细胞胚胎发生过程中下胚轴脱分化和再分化的影响,优化体细胞胚胎发生体系和初步解析脱落酸调控棉花体细胞胚胎发生分子机制。【方法】以棉花品种中棉所24(CCRI 24)下胚轴为外植体,设置5个ABA浓度0、0.02、0.04、0.06、0.08μmol·L^-1,分别以A0、A1、A2、A3、A4表示,添加至MSB(MS培养基+B5维生素)培养基诱导愈伤和胚性愈伤,研究ABA对棉花下胚轴初始细胞脱分化、愈伤组织诱导和胚性愈伤组织诱导的影响。【结果】ABA促进下胚轴初始细胞脱分化;显著提高愈伤组织的脱分化率和增殖率;0.02μmol·L^-1ABA显著提高胚性愈伤分化率,0.04~0.08μmol·L^-1ABA显著降低胚性愈伤分化率。ABA处理后胚性愈伤和非胚性愈伤的增殖率均显著提高且质地受到影响。0.02~0.08μmol ABA处理下,LBD和LBD在愈伤起始期上调表达。0.02μmol·L^-1ABA处理下,在愈伤增殖早期和中期BBM、LEC1和AGL15上调表达,愈伤增殖后期FUS3、LEA、ABI3基因上调表达。【结论】脱落酸调控的棉花体细胞胚胎发生与相关标记基因的时空性表达密切相关,这些基因表达水平的增加是ABA调控愈伤和胚性愈伤分化的分子基础。 相似文献
15.
Li Shuye Ma Huijuan Zhang Siping Liu Shaodong Shen Qian Chen Jing Ge Changwei Pang Chaoyou Zhao Xinhua 《棉花学报》2018,30(3):252-260
[Objective] The objective of this study was to investigate the effects of exogenous 24-epibrassinolide (EBR) on the photosynthetic physiology of cotton seedlings under low temperature and to provide basis for improving the cold tolerance of cotton by using EBR as growth regulator. [Method] Taking CCRI 60, Lumianyan 28 and Simian 3 as materials, a field experiment was carried out in Institute of Cotton Research of CAAS(Anyang county, Henan province). Before the first low temperature treatment, the cotton seedlings were sprayed with distilled water (Control) and different concentrations of EBR (0.1 mg·L－1 and 0.2 mg·L－1), respectively. After 3 days, the relative electrical conductivity, chlorophyll content, rapid chlorophyll fluorescence induction kinetic curve (OJIP) and fluorescence parameters were measured. [Result] Under low temperature, the relative conductivity of CCRI 60, Lumianyan 28 and Simian 3 spraying with EBR decreased by 17.7%～32.8% compared with control, and there was no significant difference between CCRI 60 and Lumianyan 28 in different concentrations of EBR treatments, butthe relative conductivity of Simian 3 treating with 0.2 mg·L－1 EBR was significantly lower than those treatments with 0.1 mg·L－1 EBR . The chl a and chl b contents increased by 9.7%～32.6% and 15.0%～18.9%, respectively. The maximum photochemical efficiency of photosystemⅡ (Fv/FM) and photosynthetic performance index on absorption basis(PIABS) increased significantly. PIABS of CCRI 60 increase the maximum by 75.6% using 0.1 mg·L－1 EBR. Lumianyan 28 and Simian 3 increased the maximum by 101.1% and 265.6% using 0.2 mg·L－1 EBR, respectively; Absorbed photon flux per cross section (ABS/CSm), electron transport flux (further than QA) per active reactive center (ETo/RC) and probability for electron transport (φEo) are significantly increased. [Conclusion] Exogenous EBR can enhance the ability of low temperature tolerance of cotton seedlings and alleviate the inhibition of photosynthesis in cotton at low temperature. The study showed that 0.1 mg·L－1 EBR performs well in CCRI 60 and 0.2 mg·L－1 in Lumianyan 28 and Simian 3. 相似文献
16.
Chen Li Liu Liantao Ma Tongtong Jiang Dan Sun Hongchun Zhang Yongjiang Zhang Ke Bai Zhiying Li Cundong 《棉花学报》2019,31(5):438-447
[Objective] Cotton (Gossypium hirsutum L.) seed germination, in which melatonin plays an important regulatory role, is seriously affected by soil salinization. Cotton seed germination, antioxidant enzyme activity levels and other physiological indicators were analyzed to clarify the regulatory effects of exogenous melatonin on cotton seed germination under salt-stress conditions. [Method] The experiment was conducted in a greenhouse of Hebei Agricultural University in Baoding City, Hebei Province from 2018 to 2019 using Guoxin Cotton 9 as the material. The germination rate, germination potential, seed biomass after germination, superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities, as well as the malondialdehyde (MDA) content, of cotton seeds were measured after exposure to different melatonin concentrations (0, 10, 20, 50 and 100 μmol·L－1) and 150 mmol·L－1 NaCl treatments. [Result] After germination, the germination rate, germination potential, radical length and seed biomass significantly decreased, as did the SOD, POD and CAT activities. However, the MDA content significantly increased under salt-stress conditions. Low exogenous melatonin concentrations (10 and 20 μmol·L－1) increased the germination potential, germination rate and biomass of cotton seeds, and promoted the elongation of radicals; however high melatonin concentrations (50 and 100 μmol·L－1) inhibited cotton seed germination and decreased seed biomass after germination. Low melatonin concentrations (10 and 20 μmol·L－1) increased the SOD, POD and CAT activities and decreased the MDA content in cotton seeds. [Conclusion] Low melatonin concentrations could promote germination and improve the salt tolerance of cotton seeds, while high melatonin concentrations could inhibit their germination. A 20 μmol·L－1 melatonin concentration is appropriate for regulating cotton seed germination under salt-stress conditions. 相似文献
17.
[Objective] Novel germplasm of cotton was generated for development of cultivars with high resistance to glyphosate. [Method] The glyphosate resistance gene G10eve was preliminarily analyzed using bioinformatics methods and introduced into 'Coker 312' via Agrobacterium-mediated transformation. The expression of G10eve was estimated by PCR (Polymerase chain reaction) and qRT-PCR (Quantitative real-time PCR). The content of shikimic acid and tolerance to glyphosate were determined in response to application of glyphosate to 'Coker 312' and transgenic plants. [Result] A total of 28 independent positive transgenic lines were obtained. The transformation efficiency and the rate of seedling differentiation were up to 49.3% and 40.6%, respectively. PCR and qRT-PCR analysis showed that G10eve was integrated into the cotton genome, and the G10eve expression level differed significantly among the transgenic lines. At the cotyledon stage, the transgenic cotton showed resistance to 8 mL·L－1 glyphosate, whereas 'Coker 312' showed phytotoxicity to the 2 mL·L－1 glyphosate treatment. No significant accumulation of shikimic acid was detected in transgenic plants before or after glyphosate treatment, which indicated the typical physical characteristic of glyphosate resistance. [Conclusion] Overexpression of G10eve in cotton improved resistance to glyphosate. Thus, novel germplasm of cotton resistant to glyphosate was generated. 相似文献