Studies on the relationship between cyanide-resistant respiration and expression of alternative oxidase in mung bean using antibodies prepared by synthetic polypeptide

Twelve peptides, including eight conservative amino acid residues in the amino acid sequence of hydrophilic S helix of the alternative oxidase (AOX), were synthesized by solid-phase method. The polypeptide was coupled with α-chymotrypsinogen, and the antibodies were obtained through immunizing domestic rabbit by injecting this complex. By using these antibodies, which were raised to immunoreact with total proteins of purified mitochondria from different organs of mung bean seedlings, we find that there are two hybridizable AOX bands in mitochondria. Their molecular weights are about 35 and 38 ku, respectively. Moreover, the respiratory parameters of hypocotyl, true leaf and cotyledon of mung bean seedlings show that true leaf has the highest total respiration (Vt), alternative pathway (AP) capacity (Valt) and the activity of AP (ρValt) among the three organs. Vt and ρValt of cotyledon ranked the second. Hypocotyl has the lowest V, and ρValt, but its Valt is higher than that of cotyledon. These result     

利用SRAP和SSR分子标记检测分析29份棉花种质遗传完整性

利用SRAP和SSR分子标记检测29份棉花种质遗传完整性。结果表明,无论每一份不同更新发芽率水平繁殖后代的种质之间,还是每一份不同繁殖世代数种质之间,其等位基因频率差异不显著,也没有检测到稀有等位基因缺失的情况。本试验表明不同更新发芽率水平和繁殖世代数差异没有对棉花这种常异花授粉作物种质遗传完整性变化产生影响。     

用合成多肽制备抗体研究绿豆抗氰呼吸与交替氧化酶表达关系

按照交替氧化酶(alternative oxidase,AOX)位于线粒体内膜外侧亲水区S螺旋的氨基酸序列,用固相法合成由12个氨基酸残基组成的多肽,将此多肽与α-牛胰凝乳蛋白酶原A相连,用作半抗原,免疫家兔制备人工合成12肽的抗体.将此抗体与来自绿豆幼苗不同器官线粒体的总蛋白分别进行Western杂交,可观察到绿豆幼苗线粒体中具有两条清晰的AOX杂交带,它们的分子量分别为35和38 ku.同时,取绿豆幼苗的子叶、真叶和下胚轴,分别测定和计算它们的呼吸参数,可以得知真叶器官组织具有最大的总呼吸(V_t)、交替途径容量(V_alt)及交替途径实际运行量(ρV_alt);其次是子叶的V_t和ρV_alt;下胚轴的V_t和ρV_alt最低,但是V_alt却高于子叶.绿豆幼苗不同器官的有关呼吸参数测定结果与AOX表达的Western分析基本一致:真叶的V_t特别是ρV_alt最高,也具有35和38 ku的AOX的杂交多肽; 其次是子叶的V_t和ρV_alt,且在子叶中,只见一条分子量为38 ku的AOX多肽;下胚轴的V_alt和ρV_alt都最低,Western杂交显示只有一条分子量为38ku的多肽,而且表达量较少.35 ku的AOX可能对真叶的ρV_alt作出了主要贡献.