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周晓婴

作品数:22 被引量:99H指数:5
供职机构:江苏省农业科学院更多>>
发文基金:江苏省农业科技自主创新基金引进国际先进农业科技计划国家油菜产业技术体系建设专项更多>>
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Effects of Seed Soaking with Uniconazole and Substrate Formula on Seedling Quality of Rapeseed被引量:3
2015年
[Objective] To cultivate blanket rapeseed seedlings suitable for automatic mechanical transplanting, the effects of seed soaking with uniconazole and substrate formula on seed germination and seedling quality of rapeseed were investigated using HY4 as the material. [Method] The rapeseed seeds were soaked in different concentrations(0, 5, 10, 20, 30, 40 and 50 mg/L) of uniconazole for 0, 1, 2, 4 and8 h, respectively. Subsequently, they were dried naturally. After the preliminary screening carried out in petri dishes and in a laboratory, the optimum soaking concentration and time of uniconazole were obtained. On the other hand, the rapeseed seeds were soaked in different concentrations(0, 5, 10, 15 and 20 mg/L) of uniconazole for 0, 1 and 2 h, respectively for outdoor pot experiment. At the same time, the screening of optimum substrate formula for cultivating blanket rapeseed seedlings was carried out. The field test was carried out to verify the screened optimum substrate formula. [Result] The results of petri dish experiment showed that seed soaking with uniconazole could delay seed germination, reduce seed germination rate, inhibit hypocotyl elongation and promote root growth. When the soaking concentration exceeded 30 mg/L and soaking time exceeded 2 h, the effect of seed soaking with uniconazole on seed germination was greater, and the seed soaking inhibited root growth. The results of outdoor pot experiment showed that the soaking concentrations and times all significantly reduced seedling height, slowed shoot growth and promoted root growth, which all contributed to the cultivation of blanket rapeseed seedlings suitable for automatic mechanical transplanting. In the substrate formula, the lower the proportion of soil was, the better the seedling growth was. In the substrate with lower proportion of soil, the seed germination rate, plant height,root vigor and dry matter accumulation were all increased(P〈0.05), which was verified by the field test. When the soaking concentration was less than 10
高建芹浦惠明龙卫华胡茂龙陈松周晓婴张维
关键词:UNICONAZOLE
甘蓝型油菜CRABS CLAW基因克隆及其RNA干扰载体的构建被引量:2
2015年
CRABS CLAW(CRC)转录因子是YABBY基因家族成员,在植物花器官发育过程中起重要作用。为进一步研究CRC转录因子在调控油菜花器官发育过程中的作用,本研究以甘蓝型油菜品种宁油10号花蕾RNA为材料,通过反转录PCR克隆到1个580 bp长度的CRC基因,并利用p Hurricane中间载体构建CRC基因的反向重复序列表达框,将CRC基因片段以正向的方式连接在1个可剪切的内含子的5'末端,以反向的方式连接到该内含子的3'末端;然后将Ca MV35S启动子序列和CRC基因的反向重复表达框再克隆到植物双元载体p CAMBIAI1390的p UC18多克隆位点,构建了干扰表达载体p A6-CRCi,经过酶切鉴定和测序分析,所构建的载体正确。
周晓婴付三雄陈松张超戚存扣
关键词:油菜RNA干扰载体
油菜抗咪唑啉酮类除草剂基因BnALS1R等位基因特异PCR标记的开发与应用被引量:13
2013年
油菜抗咪唑啉酮类除草剂基因BnALS1R是从抗性突变体M9中克隆获得,抗性基因BnALS1R与野生型基因BnALS1存在1处SNP,即乙酰乳酸合酶第638位丝氨酸残基被天冬酰胺酸替代。为获得油菜抗除草剂基因BnALS1R的分子标记,根据该处点突变,结合获得的BnALS3与BnALS1序列,开发30条等位基因特异PCR(allele-specific PCR,AS-PCR)引物,采用筛选出的3条AS-PCR引物在F2、BC1和BC2群体中进行PCR扩增。结果表明,该标记有效检测出群体中存在的3种基因型,其分离比分别为1∶2∶1、1∶1、1∶1,均遵循单基因遗传规律。应用该标记对获得的抗性恢复系进行PCR扩增,结果发现所有抗性恢复系均能扩增出抗性基因BnALS1R目的条带,表明3条标记引物可应用于抗性基因的检测。AS-PCR标记的获得将促进以抗性基因进行油菜抗除草剂分子标记辅助选择育种。
胡茂龙龙卫华高建芹付三雄陈锋周晓婴彭琦张维浦惠明戚存扣张洁夫陈松
关键词:油菜乙酰乳酸合成酶
油菜(Brassica napus L.)BnPGIP基因克隆及其表达分析被引量:2
2011年
以油菜品系甘蓝型油菜宁RS-1为植物材料,以南京油菜田里收集的核盘菌为致病菌,根据已经报道的油菜PGIP基因序列,设计简并引物。从宁RS-1基因组DNA中经PCR克隆到1条包含1个内含子与2个外显子的基因序列。序列比对分析显示,该基因与已公布的油菜PGIP1基因的核苷酸序列和氨基酸序列同源性均达99%,编码区序列全长984 bp,编码332个氨基酸,富含亮氨酸。以翻译延伸因子EF-1α作为内标,利用半定量RT-PCR分析了BnPGIP基因在核盘菌未诱导和诱导后宁RS-1中的表达量差异。结果发现,核盘菌诱导后宁RS-1中的BnPGIP表达含量明显提高,表明宁RS-1在受到核盘菌侵染后体内的BnPGIP基因受到诱导表达。为进一步研究BnPGIP基因在抗核盘菌侵染的作用,构建了BnPGIP基因过量表达载体。
周晓婴陈松戚存扣
关键词:甘蓝型油菜多聚半乳糖醛酸酶抑制蛋白基因克隆
甘蓝型杂交油菜新品种宁杂31的选育被引量:1
2016年
宁杂31是以MI CMS雄性不育系宁A7为母本、抗倒高油恢复系09N59为父本育成的双低杂交油菜新品种,具有综合性状较好的特点,具体特征为株高188.2 cm,一次分枝8.9个,单株有效角果408.4个,每角20.4粒,千粒质量3.6 g,芥酸含量0.16%,硫苷含量24.16μmol/g,含油量43.96%。宁杂31丰产稳产性好,适合在江苏省及邻近省(市)油菜主产区种植。宁杂31耐盐性较强,在江苏沿海滩涂盐碱地种植也具有较好的丰产性。
龙卫华浦惠明胡茂龙高建芹张洁夫戚存扣陈松陈锋付三雄周晓婴张维彭琦王晓东
关键词:杂交油菜选育耐盐盐碱地
核盘菌诱导下甘蓝型油菜宁RS-1的SSH文库构建被引量:3
2011年
以甘蓝型油菜宁RS-1为材料,用核盘菌(Sclerotinia sclerotiorum(Lib.)de Bary)对幼苗进行接种,提取经核盘菌诱导前和诱导后的宁RS-1叶片mRNA,经反转录成cDNA,以及经过酶解后加接头和两次杂交,构建差异表达的抑制消减(SSH)cDNA文库。正向消减文库是以诱导后的宁RS-1 cDNA为tester,以正常的宁RS-1 cD-NA为driver。所构建的正向文库重组率达到96.2%,反向文库重组率达到94.5%,文库容量均达到了2 190和2 780,表明所构建的文库质量良好。对从文库中随机挑取的克隆进行测序分析,获得了与抗病和代谢有关的基因,如γ-谷氨酰半胱氨酸酶基因、抗真菌蛋白基因、铁蛋白基因、ACC氧化酶基因等,为开展油菜抗菌核病重要功能基因的克隆和鉴定奠定了基础。
周晓婴陈松戚存扣
关键词:甘蓝型油菜核盘菌抑制消减杂交CDNA文库
Analysis of Seed-specificity of Silencing fad_2 Gene Expression in Transgenic Rapeseed Line W-4(Brassica napus L.)被引量:3
2014年
This study was to investigate the efficiency and specificity of RNAi silencing on the expression of endogenous fad2 gene in transgenic line W-4. [Method] The relative expression of fad2 gene in seeds at different developmental stages of 7th, 14th, 21st and 28th day after flowering (DAF) as wel as the root, stem, leaf at winter seedling stages of both the transgenic line W-4 and non-transgenic control Westar by real-time fluorescence quantitative PCR. [Results] The results showed the relative expression of fad2 gene was gradual y increasing with the days after flowering in the seeds of the control Westar, while it was found decreasing significantly since the 21st DAF in the seeds of the line W-4. The decline was up to 60% in comparison with the control Westar. However, no significant difference in the relative expression of fad2 gene in other organs like root, stem and leaf was observed between transgenic line W-4 and non-transgenic control Westar. Fatty acid composition analysis showed the oleic acid desaturation parameter(ODP) in seeds of the line W-4 was 0.07 in average, decreased by nearly 75% than control Westar which was 0.24 in average, while no significant difference in the seedling root, stem and leaf was measured between transgenic rapeseed and control. [Conclusion] The results above validated that RNA interference in transgenic rapeseed W-4 is at a seed-specific manner, not interfering with fad2 gene expression in organs such as the root, stem and leaf. The study also found that the period of fad2 gene expres-sion decline was wel coincided with the expression of napin gene, both appeared at the 21st DAF, indicating that the expression of dsRNA of fad2 gene is precisely control ed by the napin promoter.
陈松彭琦周晓婴高建芹张维张洁夫浦惠明戚存扣
甘蓝型油菜离体培养再生体系的优化被引量:4
2016年
为优化甘蓝型油菜离体培养再生体系,本研究分析了影响再生频率的多个关键因素,包括品种、外植体类型、苗龄、预培养时间以及激素浓度与配比等。结果表明:品种对再生频率影响较大,Westar的外植体再生频率高于宁油18号和中双11号;不同外植体再生频率也存在差异,子叶柄的再生频率显著高于下胚轴;苗龄以4 d较好,预培养时间以2 d较适宜;预培养基中激素2,4-D最适浓度为1 mg/L,分化培养基中,6-BA/NAA比例为10∶1时子叶柄的再生频率最高,为70.79%,6-BA/NAA比例为9∶1时下胚轴的再生频率最高,达到96.73%。经过优化后,油菜外植体再生频率得到显著提高。
郭婷婷周晓婴张维陈锋彭琦陈松张洁夫
关键词:油菜离体培养影响因素
宁RS-1 SSH cDNA文库构建和PGIP基因克隆及其表达载体构建
油菜(Brassica napus)是我国四大油料作物之一,菜籽油更是我国主要的食用植物油。菌核病是油菜的重要病害,它是由核盘菌(Sclerotinia sclerotiorum)侵染植株引起的茎、枝腐烂,导致油菜产量损...
周晓婴
关键词:油菜文库构建基因克隆
文献传递
Construction of a RNA Interference Vector of Brassica napus L.Pyruvate Kinase Gene
2014年
A predominantly expressed gene, pyruvate kinease (PK) gene, control ing oil accumulation, had been identified in previous study. To construct a PK RNAi vector, a 498-bp target PK gene sequence was amplified and transferred into the pEASY-T1vector. Subsequently, the target DNA fragments were cut off by enzymes Not I and Xho I and directional y inserted into plant RNAi platform vector pHurricane, a newly developed RNAi vector in our lab, to form the PK inverse repeats. The PK inverted repeats fragment was then cloned into a modified vector pCAMBIA1390, driven by a rapeseed seed-specific promoter napin. Restriction enzyme digestion verified the successful construction of RNA interference vector. The PK RNAi con-struction wil lay a foundation for study in the function of PK in oil accumulation and metabolism in rapeseeds.
付三雄张超陈松周晓婴戚存扣
关键词:RNAINTERFERENCE
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