共查询到19条相似文献，搜索用时 234 毫秒
1.
油菜高效转化系统的研究 总被引：17，自引：1，他引：16
以甘蓝型油菜湘油13为试验材料,运用子房注射法将BT毒蛋白基因导入油菜。在授粉后第20小时至第30小时用自制的微玻针从子房中部注射0.5~1.5μg外源DNA,以高达12.8%的频率获得抗卡那霉素植株。分子杂交分析证明BT毒蛋白基因已整合到油菜基因组中,转化频率为1.6%,表明子房注射法是一种有效、实用的油菜遗传转化方法。 相似文献
2.
农杆菌介导NHX基因转化甘蓝型油菜的研究 总被引：16，自引：0，他引：16
以甘蓝型油菜带柄子叶为转化受体，通过根癌农杆菌EHA105（Agrobacterium tumefaciens）介导将盐地碱蓬（Suaeda heteroptera）的Na+/H+反向运输体基因NHX导入新疆主栽油菜1khp11品系中，获得了抗卡那霉素的再生植株，并对影响遗传转化的一些关键因素进行了研究。结果表明,带柄子叶在含有2,4-D的培养基上经过3 d预培养后于浓度OD600为0.3～0.4的菌液中浸染7 min, 卡那抗性绿苗率可达8%～9%，经过对抗性植株的PCR检测、点杂交及PCR-Southern杂交证明外源基因已整合到油菜基因组中，为新疆盐碱土上油菜产量的提高提供了新的途径。 相似文献
3.
4.
5.
以番茄新品种T431、1911、0701为材料,以9天苗龄的叶片为受体,通过农杆菌Ri质粒介导,将几丁质酶基因和?-1,3葡聚糖酶基因转化番茄。共获得再生植株100株,经PCR技术检测,导入几丁质酶基因和?-1,3葡聚糖酶基因的植株分别为27株和13株,转化率为27% 和13%;两个目的基因同时导入的植株10株,转化率为10%。PCR-Southern检测呈阳性,表明外源基因已经整合到番茄基因组中。对9株双转化的植株进行RT-PCR检测,几丁质酶基因和?-1,3葡聚糖酶基因的阳性植株分别为5株和4株,证明转化的基因已表达。 相似文献
6.
8.
利用黄籽甘蓝型油菜自交系建立和优化了遗传转化系统。首先构建了由质粒pCNR与Δ6脂肪酸脱氢酶基因插入到植物的高效表达载体pCAMBIA2301G。利用在Murashige和Skoog培养基(含有200 mmol L–1乙酰丁香酮)培养5~7 d的下胚轴外植体与农杆菌株LBA4404共培养63~69 h (pCNR),再于芽诱导培养基上培养3个月诱导芽再生。在最佳条件下,平均转化效率约为1.3%。转化植株的GUS分析和PCR分析结果表明,外源基因成功导入甘蓝型油菜。Southern杂交表明,这些转化子含有目标基因1~2个拷贝。用气相色谱分析转基因植物种子的脂肪酸,γ-亚麻酸含量达8.2%。 相似文献
9.
山菠菜胆碱单加氧酶基因对棉花的遗传转化和耐盐性表达 总被引：13，自引：0，他引：13
胆碱单加氧酶（CMO）是渗透保护剂甜菜碱生物合成的关键酶之一。以棉花（Gossypium hirsutium L.）泗棉3号的下胚轴切段为外植体，利用农杆菌介导法将克隆自山菠菜（Atriplex hortensis）的AhCMO基因导入其中，通过组织培养胚状体发生途径获得转基因再生植株。以0.5%卡那霉素对再生苗筛选后，PCR检测抗卡那霉素棉苗确认阳性转化株，Southern和Northern杂交结果进一步证实外源基因已导入棉花并得到表达。在温室盆栽条件下，于2片真叶期对转AhCMO基因棉花及其转化受体泗棉3号施加0.5%的NaCl胁迫，15 d后发现其光合作用和植株生长被显著抑制，非转基因棉花泗棉3号的株高、鲜重和光合速率分别降低了57.6%、65.6%和69.9%，而转AhCMO基因的棉株分别降低了37.3%、54.6% 和47.9%。转AhCMO棉花所受盐害程度显著小于非转基因的棉株，说明AhCMO基因的导入和表达提高了转基因棉花的耐盐性。 相似文献
10.
11.
Production of fertile transgenic Brassica napus by Agrobacterium-mediated transformation of protoplasts 总被引：3，自引：0，他引：3
A protocol for Agrobacterium tumefaciens‐mediated transformation of Brassica napus mesophyll protoplasts is described. A strain with a neomycin phosphotransferase (nptII) gene and a KCS gene under control of a napin promoter was used at co‐cultivation. Transformed protoplasts were regenerated to fertile and morphologically normal transgenic plants. Transformants were confirmed by PCR of the nptII gene and NAP/KCS expression cassette, and Southern blot analysis. Seeds of the transformants showed a changed fatty acid profile: two transformants had a higher erucic acid level and differed significantly from that of B. napus. Genetic analysis of the progeny revealed that the kanamycin resistance introduced was inherited in a Mendelian fashion. 相似文献
12.
13.
M. A. Bhat M. L. Gupta S. K. Banga R. K. Raheja S. S. Banga G. Rakow 《Plant Breeding》2002,121(5):456-458
Genetic studies were undertaken to reassess erucic acid heredity in Brassica juncea. Analysis of segregation in F2 and BC1 generations from two zero × high erucic acid crosses indicated that higher erucic acid in B. juncea was controlled by two dominant genes with additive effects, whereas segregation in a cross involving ‘CCWF 16′, a genotype having intermediate erucic acid (25.6%), and a zero erucic acid strain, indicated monogenic dominant control for intermediate erucic acid content. The B. juncea strain ‘CCWF 16’ was developed by hybridizing high‐erucic acid B. juncea cv.‘WF‐1’ with a ‘0’ erucic B. rapa cv.‘Candle’ followed by backcrossing with ‘WF‐1’ and half‐seed selection for low erucic acid in each backcross generation. This strategy resulted in substitution of the high erucic acid allele present in the A genome of B. juncea (AABB) by the zero erucic acid allele associated with ‘A’ genome of ‘Candle’. The intermediate erucic acid content in ‘CCWF 16’ was thus attributed to a gene present in the ‘BB’ genome. Experimental data clearly suggested that the gene (E2) associated with the A genome had a greater contribution to the total erucic acid content in B. juncea than the gene (E1) located on the B genome. This provided experimental evidence for a previous suggestion of unequal contributions of two dominant genes (E1= 12%, E2= 20%) to high erucic acid content in conventional digenomic Brassica species. 相似文献
14.
Independent Inheritance of Erucic Acid Content and Flower Colour in the C-Genome of Brassica napus L. 总被引：3，自引：1，他引：3
The synthetic Brassica napus L. line No7076 was obtained from a cross between yellow-flowered and zero-erucic turnip rape (B. campestris) Sv85-38301 and white-flowered and high-erucic (41.4%) B. oleracea ssp. alboglabra No6510. This synthetic B. napus is pale-flowered and has an average erucic acid content of 25.8 %. It was crossed with the yellow-flowered and zero-erucic B. napus line SvS4-2S053 and segregation of the erucic acid content and flower colour was studied in F1 and F2 generations. The high erucic acid content was controlled by a single gene in the C-genome and was additively inherited. Strong evidence was obtained in support of independent segregation of the erucic-arid content and the flower colour characters controlled by the C-genome of B. napus. 相似文献
15.
Two yellow‐seeded white‐petalled Brassica napus F7 inbred lines, developed from interspecific crosses, containing 26–28% emcic acid and more than 40 μmol glucosinolates (GLS)/g seed were crossed with two black/dark brown seeded B. napus varieties of double low quality and 287 doubled haploid (DH) lines were produced. The segregation in the DH lines indicated that three to four gene loci are involved in the determination of seed colour, and yellow seeds are formed when all alleles in all loci are in the homozygous recessive state. A dominant gene governed white petal colour and is linked with an erucic acid allele that, in the homozygous condition, produces 26–28% erucic acid. Four gene loci are involved in the control of total GLS content where low GLS was due to the presence of recessive alleles in the homozygous condition in all loci. From the DH breeding population a yellow‐seeded, yellow‐petalled, zero erucic acid line was obtained. This line was further crossed with conventional B. napus varieties of double low quality and, following pedigree selection, a yellow seeded B. napus of double low quality was obtained. The yellow seeds had higher oil plus protein content and lower fibre content than black seeds. A reduction of the concentration of chromogenic substances was found in the transparent seed coat of the yellow‐seeded B. napus. 相似文献
16.
ABI4(Abscisicacid-insensitive4)是ABA响应途径中重要的转录因子。本研究以甘蓝型油菜全基因组数据库为基础,借助拟南芥ABI4蛋白的氨基酸序列,通过序列比对,获得了甘蓝型油菜的2个BnaABI4,分别命名为BnaABI4-A (GenBank登录号为BnaA03g18970D)和BnaABI4-C (GenBank登录号为BnaC03g725-10D)。在这两个基因的CDS序列上寻找一段长度为20 bp、序列相同的片段作为基因编辑的目标位点,构建CRISPR/Cas9基因编辑载体pCAMBIA1300-sg RNA/Cas9-BnaABI4;借助根癌农杆菌介导的油菜下胚轴遗传转化法,成功将BnaABI4的基因编辑表达框转入甘蓝型油菜‘湘油15’中,共获得转基因植株13株,这将为深入研究甘蓝型油菜BnaABI4的生物学功能提供科学依据。 相似文献
17.
甘蓝型油菜DH群体主要品质性状相关性及主成分分析 总被引：4，自引：3，他引：1
为了研究甘蓝型油菜品质性状之间的相关性,以期有效的改良油菜的品质性状,将170份DH纯系群体于2009-2012年进行田间试验,检测油菜种子品质性状,对其表型、相关性及主成分进行分析。结果表明：亲本中芥酸含量的变异系数最大(相差15.53%)。油菜DH群体中硫甙含量的变异幅度最大(128.43μmol/g饼粕),饱和脂肪酸的变异幅度最小(1.97%)。DH群体的品质性状表型变异幅度显示出了超亲遗传的现象。除了芥酸,含油量与其他组分呈现出极显著的负相关,特别是与蛋白质含量的负相关系数最大(-0.72)；蛋白质含量与油酸、亚油酸、亚麻酸和硫甙含量均呈极显著的正相关,与含油量和芥酸呈极显著的负相关；除了蛋白质,其他组分含量皆与硫甙含量呈现出极显著或者不显著的负相关。通过主成分分析可将品质性状综合为油酸亚油酸酸因子,蛋白质因子,亚麻酸因子和硫甙因子,累积贡献率达到93.94%,基本反映甘蓝型油菜DH系群体脂肪酸组分信息。 相似文献
18.
19.
A greenhouse study was conducted to determine the effect of nitrogen supply (30, 100 or 170 ppm N) and raceme position on the fatty acid composition of oil extracted from erucic acid-free summer rape seed ( Brassica napus cv. Callypso ). The seven fatty acids analyzed for include palmitic, palmitolcic, stearic, oleic, linoleic, linolemc, and eicosenoic acids; of which oleic (59.54–64.84 %) and palmitoleic (0.36–0.4 %) acids were the highest and lowest levels respectively. Generally, N nutrition influenced fatty acid pattern only to a little extent. Palmitic, palmitoleic and stearic acid levels were increased by 170 ppm N, depending on raceme position, but oleic and linolenic acids were unaffected. Similarly, 170 ppm N produced the highest fatty acid levels in seeds on the lower portions of racemes, with the exception of oleic acid. This was also true in the case of the upper portions of racemes, except that 30 ppm N produced the highest levels of oleic and linoleic acids in rape seeds. Under the optimum N supply level (i.e. 100 ppm N), position of raceme on the rape plant did not greatly influence the levels of different fatty acids in lipids. 相似文献