低钙敏感拟南芥自然变异种的筛选及相关基因的粗定位生物工程专业论文.docxVIP

内蒙^人学倾Jj学化论义WITH 内蒙^人学倾Jj学化论义 WITH LOW CALCIUM SENSITIVE PHENOTYPE 0F ARABIDOPSIS THALIANA NATURAL MUTANT SELECTl0N AND ROUGH NIAPPING OF RELATED GENE ABSTRACT In order to understand the molecular mechanism of the plant growth in the low calcium situation，this experiment mainly makes the natural mutant Arabidopsis thaliana as material，filtrates out a kind of low calcium sensitive natural mutant Wt一5 through the filter of 96 kinds of natural variation types．By the phenotypic analysis，it is found that compared with Arabidopsis wild—type Col一0，Wt-5 has the phenotype of short roots and small and yellow leaves under the situation of the low calcium medium(contains 09M CaCl2，20pM CaCIz)，which ahnost has no differences with the phenotype in CK medium(contains l lnM CaCl2)． By atomic absorption spectrum method to measure the total calcium of Wt一5，it is found that compared with wild type Col一0．the total calcium content of Wt_5 grown in the CK is lower．Receiving low calcium treatment，Wt一5 still remains a lower content of the total calcium in the leaf(1arge leaf or small leaf)． F I and F2 plants can be got by hybriding Wt一5 and Ler(ecological)，which has been shown to be the single recessive mutant genes by analysis．Using the map—based 第2页 万方数据 内蒙古人学坝上学位论文cloning．mutations 内蒙古人学坝上学位论文 cloning．mutations can be roughlY positioned within the scope of 1 8．14M一19．5Mbp of Arabidopsis chromosome 2． KEYWORDS：Arabidopsis；Natural variation；Low calcium；Phenotypic analysis； M印-based cloning technology 第3页 万方数据 内蒙占人学硕上学位论文目录 内蒙占人学硕上学位论文 目录 符号说明 一1 一引言 ．2 1．1钙离子对植物的生理作用 2 1．2植物吸收钙离子的分子机制 ．2 1．2．1钙调素及类钙调素蛋白(CaM) ．3 1．2．2钙调磷酸酶B类蛋白(CBL) ．4 1．2．3钙依赖性蛋自激酶(CDPK) 5 1．3钙资源现状 7 1．3．1酸雨对土壤中钙的淋失 ．．7 1．3．2酸雨对植物中钙的影响 ．．8 1．4图位克隆的基本原理 一8 1．5本研究的目的与意义 ．．9 二材料与方法 ．．1 0 2．1材料 一1 0 2．1．1植物材料 ．10 2．1．2主要试剂 lO 2．1．3主要溶液与培养基的配制 10 2．1．4引物列表 ．11 2．2方法 14 2．2．1拟南芥的播种与培养 14 2．2．2自然变异种Wt一5的筛选 一1 5 2．2．3 Wt一5水培表型观察 一1 5 2．2．4原子吸收测定总钙含量 1 5 2．2．5 CTAB法提取DNA ．．1 6 22．6 Wt一5相关基因定位(图位克隆) 第4页 万方数据 内蒙_^J大学硕上学位论文三结果与分析 内蒙_^