大白菜突变体库的构建及M₂叶片表型变异的研究     

卢 银刘梦洋  赵建军王彦华罗双霞  轩淑欣代双燕王超硕  申书兴 《园艺学报》2014,41(8):1609-1619

 利用不同浓度甲基磺酸乙酯（EMS）诱变大白菜自交系‘A03’种子,根据M₁代植株发芽率、成活率、育性及M₂代成活率,筛选出适宜的EMS诱变浓度为0.4%。采取EMS种子诱变1次、EMS种子诱变2次及EMS花蕾诱变结合小孢子培养的方法构建了含有4 253个家系及相应自交M₂代种子的大白菜突变体库。在该突变体库M₂群体中分别对苗期6 404株幼苗及莲座期7 756个单株的叶片性状进行了形态学调查,总的表型变异频率高达37.62%和26.33%。  相似文献   

Sensitive detection of the c‐KIT c.1430G>T mutation by mutant‐specific polymerase chain reaction in feline mast cell tumours     

M. Takanosu  M. Sato  Y. Kagawa 《Veterinary and comparative oncology》2014,12(2):138-142

Here, we describe the establishment of mutant‐specific polymerase chain reaction (PCR) for detection of a c‐KIT c.1430G>T mutation in feline mast cell tumours. Several mutations in feline c‐KIT have been identified, with the c.1430G>T mutation accounting for a significant portion of feline mast cell tumour mutations. The c.1430G>T mutation in c‐KIT exon 9 was detected in 15.7% (11 of 70) of samples by mutant‐specific PCR but in only 7.1% (5 of 70) by PCR–restriction fragment length polymorphism (RFLP) in the genomic DNA isolated from 70 formalin‐fixed paraffin‐embedded sections or cells collected by fine needle aspiration. Mutant‐specific PCR showed remarkably higher detection rate than did PCR–RFLP. DNA sequence analysis did not always yield identical results to those of mutant‐specific PCR, suggesting heterogeneity of tumour cells. Mutant‐specific PCR is a valid and efficient screening tool for detection of the c‐KIT c.1430G>T point mutation in feline mast cell tumours compared with PCR–RFLP and sequencing analysis.  相似文献   

Use of advanced recombinant lines to study the impact and potential of mutations affecting starch synthesis in barley     

Thomas P. Howard  Brendan Fahy  Fiona Leigh  Phil Howell  Wayne Powell  Andy Greenland  Kay Trafford  Alison M. Smith 《Journal of Cereal Science》2014

The effects on barley starch and grain properties of four starch synthesis mutations were studied during the introgression of the mutations from diverse backgrounds into an elite variety. The lys5f (ADPglucose transporter), wax (granule-bound starch synthase), isa1 (debranching enzyme isoamylase 1) and sex6 (starch synthase IIa) mutations were introgressed into NFC Tipple to give mutant and wild-type BC₂F₄ families with different genomic contributions of the donor parent. Comparison of starch and grain properties between the donor parents, the BC₂F₄ families and NFC Tipple allowed the effects of the mutations to be distinguished from genetic background effects. The wax and sex6 mutations had marked effects on starch properties regardless of genetic background. The sex6 mutation conditioned low grain weight and starch content, but the wax mutation did not. The lys5 mutation conditioned low grain weight and starch content, but exceptionally high β-glucan contents. The isa1 mutation promotes synthesis of soluble α-glucan (phytoglycogen). Its introgression into NFC Tipple increased grain weight and total α-glucan content relative to the donor parent, but reduced the ratio of phytoglycogen to starch. This study shows that introgression of mutations into a common, commercial background provides new insights that could not be gained from the donor parent.  相似文献   

Physicochemical Property Analysis and Gene Mapping of a Floury Endosperm Mutant flo7 in Rice     

FANG Peng fei  LI San feng  JIAO Gui ai  XIE Li hong  HU Pei song  WEI Xiang jin  TANG Shao qing 《中国水稻科学》2014,28(5):447

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水稻半矮杆突变体‘双辐矮糯’的农艺性状与遗传分析     

张林金  袁定阳  刘桃李  谭颖  杨震  曾建光  王学华  谭炎宁 《热带作物学报》2022,43(6):1122-1128

倒伏是影响水稻（Oryza sativa L.）生产的重要因素,半矮杆是协调高产和抗倒性的理想资源。前期通过辐射诱变‘湘辐糯1号’获得了半矮杆突变体‘双辐矮糯’。本研究分析了‘双辐矮糯’的茎节配置特点、基本农艺性状及其遗传规律。结果表明：‘双辐矮糯’株高较‘湘辐糯1号’（133.13 cm）降低了24.88 cm,穗、第1节和第2节长度显著缩短,降幅分别达到18.10%、23.06%和19.13%,而第3、4、5节长度缩短程度不明显,表明其株高突变基因主要控制穗和高位茎节的伸长。‘双辐矮糯’的单株有效穗为10.4个,是‘湘辐糯1号’（5.2个）的2.04倍,其强分蘖能力确保了在每穗总粒数下降20.24%和千粒重下降7.51%情况下单株产量仍表现出25.26%的增长。品质分析发现,‘双辐矮糯’的粒长、粒宽、长宽比等外观品质指标及直链淀粉含量、胶稠度等食味品质指标与‘湘辐糯1号’无显著差异,说明‘双辐矮糯’株高的下降未对外观品质与食味品质产生不利影响。遗传分析发现,48对SSR标记在‘双辐矮糯’与‘湘辐糯1号’间扩增出了完全一致的基因型,二者遗传背景相似度达到了100.00%,证实了‘双辐矮糯’是源于‘湘辐糯1号’的真实变异。利用‘双辐矮糯’与‘湘辐糯1号’构建了2个正反交F₂群体,分析发现群体中半矮杆植株与野生型植株理论分离比符合1∶3,表明半矮杆性状受一个隐性细胞核基因控制。本研究明确了‘双辐矮糯’的株高表型和遗传特点,为深入挖掘其育种价值奠定基础。  相似文献   

大豆脂肪酸脱氢酶GmFAD3C-1基因的克隆及功能分析     

焦苏淇  周俊名  尚雨晴  王佳欣  张爱晶  何浩博  赵秋竹  李越  姚丹 《中国油料作物学报》2022,44(5):1006

研究通过对大豆脂肪酸脱氢酶GmFAD3家族中4个关键酶基因序列进行比对,与对照JN18相比,大豆低亚麻酸突变体MT72中GmFAD3C-1基因在起始密码子后+966bp处存在一个碱基位点的缺失（G→?）,产生移码突变,导致氨基酸序列上产生较大变化（该突变基因命名为gmfad3c-1）。构建GmFAD3C-1基因超表达及CRISPR/Cas9编辑载体,利用花粉管通道法获得T₂转化植株。脂肪酸脱氢酶酶活测定结果显示,超表达植株T₂籽粒中,酶活与对照相比上升47.62%~78.85%,编辑载体T₂籽粒中,酶活与对照相比下降25.67%~47.11%。脂肪酸相对含量测定的结果进一步表明,GmFAD3C-1基因的表达与植株亚麻酸含量密切相关。  相似文献   

人泛素结合酶UbcH5c活性突变体的构建·纯化以及催化活性研究     

张西轩  李晔  阮海华 《安徽农业科学》2014,(3):663-666

[目的]构建人泛素结合酶UbcH5c的活性位点突变体S22R和F62A,同时分析这2种突变体蛋白与野生型蛋白在泛素化反应中的活性差异。[方法]通过点突变(Site-Directed Mutagenesis)的方法构建2种突变体质粒,利用0.5 mmol/L IPTG分别诱导2种突变体蛋白在大肠杆菌中进行表达;将菌体超声破碎后,依次利用阳离子交换层析法对UbcH5c突变蛋白进行分离纯化,最后利用体外泛素化酶促反应结合蛋白免疫印迹杂交的方法分析野生型UbcH5c与其活性突变体UbcH5c S22R和UbcH5c F62A在泛素化修饰上的差异。[结果]人泛素结合酶UbcH5c的2个突变体蛋白S22R、F62A的泛素化修饰程度明显弱于野生型蛋白。[结论]突变体S22R和F62A突变均不同程度的改变了人泛素结合酶UbcH5c与泛素分子之间的结合活性,即2个突变的位点中第62位苯丙氨酸残基及第22位丝氨酸残基均是UbcH5c结合泛素的关键位点,而且后者对于UbcH5c结合泛素活性的影响更大。  相似文献   

优质水稻黄华占雄性不育突变体的筛选及初步分析     

陈竹锋  卢嘉威  卢启清  王娜  王成旭  谢刚  周向阳  唐晓艳 《广东农业科学》2014,41(19):1-4

利用EMS(甲磺酸乙酯)处理优质水稻品种黄华占种子,经过田间筛选和对M1、M2代连续的遗传分析得到317份独立的、能够稳定遗传的、由单隐性核基因控制的雄性不育突变材料.通过对花器官形态观察及花粉染色分析,初步将这些突变体材料分为6类,其中花粉发育突变体在不同的花粉发育时期出现异常.这些突变材料为深入研究水稻雄性不育分子机制及开展分子设计育种提供充足的材料.98个无粉型雄性不育突变体用于雄性不育基因TDR的测序分析,其中3个突变体在3个不同的位置上出现单碱基突变.  相似文献   

Cloning and Expression of Gene Responsible for High-Tillering Dwarf Phenotype in Indica Rice Mutant gsor23     

YUAN Shou-jiang  WANG Tao  YIN Liang  ZHAO Jin-feng  WAN Jian-min  LI Xue-yong 《水稻科学》2013,20(5):320-328

High-tillering dwarf mutant gsor23 was generated from an indica rice variety Indica9 radiatied by γ-ray. Genetic analysis showed that this phenotype was controlled by one single recessive gene, which was mapped within a physical distance of 386 kb between two insertion-deletion (InDel) markers C1-WT2 and C1-WT4 on the long arm of chromosome 1. There is a known gene D10 within this region, the mutation of which causes high-tillering in rice. Sequence analysis of the D10 allele in gsor23 revealed that the base cytosine (C) at the 404^th position in the coding region was deleted, which would cause frameshift mutation after the 134^th amino acids. The mutation site and indica background of gsor23 were different from the previously reported japonica mutants d10-1 and d10-2. Therefore, gsor23 is a novel allelic mutant of D10 which encodes the carotenoid-cleaving dioxygenase 8 (CCD8), a key enzyme involved in the biosynthesis of the new plant hormone strigolactones (SLs). After treatment with GR24, a synthetic analogue of SLs, the high-tillering phenotype of gsor23 was restored to normal. Real-time RT-PCR analysis showed that D10 expression was high in roots, but low in leaves. Compared with the wild type Indica9, the expression of the SL biosynthesis gene D10 was upregulated, while genes likely involved in the SL signal transduction pathway such as D3 and D14 were down-regulated in the gsor23 mutant.  相似文献   

NYH-1型~(14)C核素高灵敏探测装置   总被引：1，自引：0，他引：1  

曲哲  时国庆  肖京城 《核农学报》2000,14(4):230-233

本文描述了可测量低活度1 4 C核素的高灵敏探测装置。在FH 36 7探头的基础上进行了改装 ,更换了闪烁介质 ,增加了物质屏蔽和反符合屏蔽以及相应的电子线路。本装置对1 4 C的探测效率 ,植物干样为 6 93% ,土壤样为 3 2 % ,装置的本底计数为 2 35cpm  相似文献