甘蓝MLPK基因的克隆与序列分析   总被引：2，自引：0，他引：2  

赵永斌  朱利泉  王小佳 《作物学报》2006,32(1):46-50

采用PCR、RT-PCR以及其他分子生物学方法，以甘蓝基因组DNA和柱头cDNA为模板对MLPK基因进行扩增克隆，首次得到长度分别为1615bp和1294bp的基因片段。序列分析表明，甘蓝MLPK与芜菁MLPK的cDNA序列相似性达98％，前者的内含子数为4个，比后者少了3个；同时，在前者内含子中发现了不同于典型的GU—AG规则的新的序列，即第3、4个内含子5，端碱基是TA、CG，3′端碱基为CAGG、GT，推测甘蓝MLPK的mRNA具有独特的剪切机制。这为甘蓝自交不亲和分子机理研究提供了新内容。  相似文献   

利用酵母双杂交法检测甘蓝SCR与SRK之间的相互作用   总被引：3，自引：1，他引：2  

罗兵  薛丽琰  朱利泉  张贺翠  彭一波  陈松  杨红  杨昆  李成琼  王小佳 《作物学报》2011,37(4):579-586

为深入研究甘蓝S位点受体激酶(SRK)和S位点富含半胱氨酸蛋白(SCR)间相互识别的分子机理,以具有典型自交不亲和性的甘蓝D3为材料,利用巢式PCR分别扩增SRK胞外域(eSRK)和SCR,通过酵母双杂交检测二者之间的相互作用。并利用同源重组技术将eSRK与GAL4报告基因DNA活化域融合(pGADT7eSRK)以及SCR与GAL4报告基因DNA结合域融合(pGBKT7SCR)。两种重组质粒分别转化酵母Y187和Y2HGold后未出现自激活现象。融合的二倍体酵母(pGADT7eSRK× pGBKT7SCR)能在选择性固体培养基(SD/–Ade/–His/–Leu/–Trp/X-a-Gal/AbA)上生长,并且菌落呈蓝色。结果表明, eSRK与SCR蛋白能够相互结合, 为深入研究二者作用位点成功建立了一个技术平台。  相似文献   

利用酵母双杂交系统鉴定羽衣甘蓝臂重复蛋白(ARC1)与结球甘蓝S位点受体激酶(SRK)激酶结构域的相互作用   总被引：2，自引：1，他引：1  

张贺翠  杨昆  朱利泉  杨永军  薛丽琰  杨红  常登龙  高启国  任雪松  李成琼  王小佳 《农业生物技术学报》2011,(6)

S位点受体激酶(SRK)和臂重复蛋白(ARC1)是芸薹属植物自交不亲和反应的两种重要的信号元件,SRK-ARC1之间的互作决定着自交不亲和反应的进程。本研究利用PCR技术获得结球甘蓝(Brassicaoleracea var.capitata)SRK激酶结构域(SRKJ)的编码区以及羽衣甘蓝(B.oleracea var.acephala)ARC1基因不同片段长度的基因组DNA与编码区,分别构建以pGBKT7为载体的长度依次递减的ARC1a、ARC1b、ARC1c和ARC1d的重组诱饵质粒和以pGADT7为载体的SRKJ的重组猎物质粒,并将重组质粒转化酵母菌株进行蛋白质相互作用检测。结果显示:羽衣甘蓝ARC1存在5个连续的臂重复区,与结球甘蓝ARC1的氨基酸序列一致性达到98%;构建的重组表达载体在酵母细胞中无毒性和自激活作用产生;3个实验组Y2HGold[pGBKT7-ARC1a]×Y187[pGADT7-SRKJ]、Y2HGold[pGBKT7-ARC1c]×Y187[pGADT7-SRKJ]和Y2HGold[pGBKT7-ARC1d]×Y187[pGADT7-SRKJ]同时激活了4种报告基因AUR1-C...  相似文献   

大白菜花粉壁蛋白和柱头表膜蛋白与自交不亲和的关系   总被引：5，自引：0，他引：5  

董耿  韦顺恋  李曙轩 《园艺学报》1993,(4)

以6个纯合S基因型(SaSa,SdSd,SfSf, SgSg,SiSi,SjSj)和1个杂合S基因型(SgSi)的大白菜自交不亲和株系为材料,提取花粉壁蛋白和柱头表膜蛋白。用花粉壁蛋白处理相同S基因型的柱头,可诱导乳突细胞的胼胝质反应:处理不同S基因型的柱头,可“蒙导”自交亲和。经SiSi柱头表膜蛋白处理过的SiSi花粉,授于花期的SjSj和SaSa以及蕾期的SiSi柱头上,原来的亲和性消失。显然,花粉壁蛋白和柱头表膜蛋白分别是花粉侧和柱头侧不亲和反应的识别物质。用等电聚焦法分析花粉壁蛋白和柱头表膜蛋白中的糖蛋白,结果表明,其等电点因S基因不同而特异。由此进一步推测,花粉壁蛋白和柱头表膜蛋白中的识别物质为S基因特异糖蛋白。  相似文献   

大白菜自交不亲和系种子生长发育规律研究     

孟祥栋  李吉奎  傅静尘 《种子》1993,(2)

以小麻叶自交不亲和系为试材,研究了大白菜种子生长发育规律。种荚长、直径授粉22天达最大,种子直径授粉29天达最大;授粉43天种荚、种子干重达最重。授粉47天,种子发芽率、活力指数最高。  相似文献   

苜蓿自交不亲和性研究   总被引：4，自引：0，他引：4  

何咏松  吴仁润 《草业科学》1987,(4)

实验中探讨了苜蓿自交不亲和性和遗传特征。结果得出:1.苜蓿自交结荚百分率为0～92％,平均为27.6％。苜蓿自交不亲和性在群体中呈连续、偏态分布。2.苜蓿自交不亲和性反应主要发生在雌蕊组织下部(子房及子房腔)。3.确定自交不亲和或自交亲和性的判别函数方程为Y=-0.0804x_1—0.5228x_2+0.6117_3。4.苜蓿自交不亲和性受单位点、复等位基因控制,并具有CSI遗传特点。5.S基因与过氧化物同工酶基因(Prxl)为连锁,两者间重组值为27％。6.PSC的遗传力h~2=62％,选择无论对亲和性和不亲和性均为有效。根据本试验结果讨论了苜蓿SI的测度指标、授粉方式及不亲和性的数量差异问题,并探讨了紫花苜蓿与黄花苜蓿种的分类差异,认为两者理应属同一种或种内不同生态型。  相似文献   

8个中国梨品种S基因型的分子鉴定     

梁文杰  谭晓风  张琳  曾艳玲  姜傲芳  曹玉芬  李秀根 《果树学报》2007,24(6):757-760

为鉴定8个中国梨品种的S基因型,使用梨自交不亲和基因(S-RNase)特异引物"FTQQYQ"和"anti-ⅡWP-NV",对8个梨品种的基因组DNA进行特异扩增,并对扩增片段进行回收、克隆、测序。使用生物信息学软件对各序列分析和经同源性搜索分析后,确定了各品种的S基因型。结果分别是:兰州花长把为S19S22,青面为S1S18,黄面为S1S12,早蜜为S19S29,大面黄为S1S19,无子黄为S28S16,大青皮为S34S19及金锤子为S16S19。  相似文献   

Heterosis of Double Low Self-incompatibility in Oilseed Rape (Brassica napus L.)     

SHEN Jin-xiong  FU Ting-dong  YANG Guang-sheng 《中国农业科学(英文版)》2002,1(7):732-737

66 F1 hybrids, produced by 3 double low self-incompatible lines and 22 varieties with a North Carolina Ⅱ (NCⅡ) crossing design, were tested for their heterosis in Wuhan, China during two growing seasons from 1999- 2001. The results showed that significant differences were found between F1s and their parents for yield per plant and seed oil content. Mid-parent heterosis of these two characters ranged from 5.50%-64.11% and from 1.55% -7.44% respectively. Heterosis for seed yield per plant was greater than that of seed oil content. For yield components, heterosis of total number of siliques per plant was the highest, followed by seed number per silique and 1 000 seeds weight. Significant genotype-by-year interaction was found for seed yield per plant. Results from correlation and combining ability analysis indicated that parental effects on its F1 hybrid depended on characters, seed yield per plant was affected by both additive and non-additive effects, and seed oil content was affected mainly by additive effect. When designing hybrid programme, parents might be selected by GCAs and variances of SCAs of parents for the characters affected by both additive and non-additive effects, and by the sum of GCAs of female and male parents for the characters mainly affected by additive effects.  相似文献   

自交不亲和植物自交亲和性研究进展     

白生文  范惠玲 《安徽农业科学》2009,37(9):3896-3897

对高等植物自交亲和基因（资源）、自交亲和性的遗传、自交亲和性与蛋白表达的关系及自交亲和性产生的分子机理等方面的研究进行了综述,阐述了研究植物自交亲和性的意义。  相似文献   

Genetic analysis reveals a dominant <Emphasis Type="Italic">S</Emphasis> locus and an <Emphasis Type="Italic">S</Emphasis> suppressor locus in natural self-compatible <Emphasis Type="Italic">Brassica napus</Emphasis>     

Chaozhi Ma  Chunyan Li  Yongqiang Tan  Wei Tang  Jianfeng Zhang  Changbin Gao  Tingdong Fu 《Euphytica》2009,166(1):123-129

A self-incompatible (SI) line, S-1300, and its maintainer 97-wen135, a self-compatible (SC) line, were used to study the inheritance of maintenance for self-incompatibility in B. napus. The ratio of SI plants to SC plants from S-1300 × 97-wen135 F₂ and (S-1300 × 97-wen135) × 97-wen135 was 346:260 and 249:232, fitting the expected ratio of 9:7 and 1:1, respectively. Based on these observations, here we propose a genetic model in which two independent loci, S locus and S suppressor locus (sp), are predicted to control the inheritance of maintenance for self-incompatibility in B. napus. The genotypes of S-1300 and 97-wen135 are S ₁₃₀₀ S ₁₃₀₀ sp ₁₃₀₀ sp ₁₃₀₀ and S ₁₃₅ S ₁₃₅ sp ₁₃₅ sp ₁₃₅ , respectively. S ₁₃₅ is dominant to S ₁₃₀₀ , but coexistence of sp ₁₃₀₀ and sp ₁₃₅ fails to suppress S locus. Both S ₁₃₀₀ and S ₁₃₅ can be suppressed by sp ₁₃₅ , while sp ₁₃₀₀ can suppress S ₁₃₅ but not S ₁₃₀₀ . The model contains two characteristics: that a dominant S locus exists in self-compatible B. napus, and that co-suppression will occur when sp loci are heterozygous. The model has been validated by the segregation of S phenotypes in the (S-1300 × 97-wen135) × S-1300, the progenies of SC S-1300 × 97-wen135 F₂ plants and DH population developed from S-1300 × 97-wen135 F₁. This is the first study to report co-suppression of S suppressor loci in B. napus. The genetic model will be very useful for developing molecular markers linked to maintenance for self-incompatibility and for dissecting the mechanism of SI/SC in B. napus.  相似文献