【作者】 许晓岗；

【导师】 汤庚国；

【作者基本信息】 南京林业大学 ， 园林植物与观赏园艺， 2006， 博士

【摘要】 本文采用IBA、NAA等外源激素，以不同浓度、不同浸泡时间处理垂丝海棠、楸子插穗。利用间接酶联免疫吸附测定法(ELISA)和聚丙烯酰胺凝胶电泳(SDS-PAGE)技术测定了不同生根型的海棠插穗生根过程中几种内源激素(TAA、ABA、ZR、GA等)和可溶性蛋白的变化。弄清了海棠扦插生根过程中，内源激素与可溶性蛋白质之间的关系。 本文还研究了海棠的内在遗传因子通过蛋白质表达来控制扦插生根过程，发现了在扦插生根过程中影响生根的可溶性蛋白种类，内外源激素通过蛋白质受体对海棠插穗生根的关系。通过3年多的研究，本文取得了以下研究成果： 1．对外源激素与扦插生根的关系的研究表明：在供试验的三种激素中，IAA具显著作用，赤霉素在结果枝的扦插中与生长素交互起显著作用。 2．不同时期扦插对插穗成活率有极显著影响。在芽萌动至展叶期，随着时间推移，扦插生根率提高，外源激素处理随时间推移效应减小。 3．研究发现，插穗的含水量、蛋白质含量、可溶性糖含量等内部生理生化因子变化对海棠扦插生根有显著的影响。通过相关系数分析，海棠扦插生根率与束缚水含量达到极显著负相关，与自由水含量显著正相关。生长促进类植物内源激素随侧芽位置而变化，离芽越远含量越低，脱落酸则出现一定的波动。扦插后含水量下降到第九天再上升，蛋白质含量、可溶性糖含量及多酚氧化酶等也缓慢上升。激素变化则表现为早期生长素／脱落酸比值及生长素／细胞分裂素类比值都在第4～6天出现峰值，早期大量生长素的产生证明是扦插所必须的。 4．对这两种海棠扦插生根过程的解剖学研究发现，垂丝海棠与楸子的插穗茎内无潜伏根原基，不定根由诱生根原基发育形成，诱生根原基源于初生射线与维管形成层交汇处细胞的分裂分化。经15天后，不定根原基发育为幼小不定根并伸出周皮之外。垂丝海棠插穗经过了愈伤组织形成、分化等过程。扦插后10天，愈伤组织开始发生；随着愈伤组织进一步分裂、分化，约至第40天愈伤组织长出不定根来。 5．对可溶性蛋白质与扦插生根研究表明： 垂丝海棠(Malus halliana Roehne)的插穗在生根过程中切口处的46KD，39KD，36KD，28KD，26KD，24KD等可溶性蛋白与插穗生根有关；39KD，26KD，24KD是三种调控蛋白，其作用是诱导愈伤组织及不定根的分化，当不定根形成后即消失。28KD和31KD蛋白是随着愈伤组织及不定根产生而新成的蛋白并促进垂丝海棠插穗的生根。46KD是一种阻碍愈伤组织再分化的蛋白，对插穗生根有抑制作用。 楸子(Malus prunifolia Borkh)的插穗在生根过程中切口处的65KD，52KD，43KD，39KD，31KD，28KD，26KD，22KD等蛋白与插穗生根有关；65KD，39KD，26KD，22KD是四种调控蛋白，其作用是诱导插穗中形成层产生根原基和不定根的分化，当不定根形成后则急剧减少或消失。52KD是一种对根原基产生和不定根的分化有阻碍作用的蛋白，对插穗生根有阻止和抑制作用。用IAA+IBA对楸子插穗处理能诱导产生72KD，83KD二种新的蛋白，这两种蛋白对插穗生根的数量及质量有强烈的促进作用。更多还原

【Abstract】 Under various treatments of exogenous phytohormone (such as IBA,NAA) in different concentration and soaking time, this dissertation tested the variances of some endogenous phytohormone (IAA,ABA,ZR,GA, etc.) and soluble proteins in cuttings with the techniques of ELISA and SDS-PAGE during the different Crab apple. cuttings’ rooting process, figured out the relations among their endogenous phytohormones and soluble proteins as well.This paper also researched the rooting process of Crab apple cuttings’ in which the inner genetic factors controlled by the way of proteins’ conveyance, discovered the kinds of proteins which affected rooting during the cutting process. It also explored the connection that endogenous and exogenous phytohormones affected rooting of Crab apple cuttings.by combining BP. Through more than three years research work, some achievements have been obtained as follow:1.Exogenous phytohormone also effected the rooting rate of cutting. Variance analysis showed that IAA had significant function on cutting. And Gibberellin mixed with IAA showed to promote each other on breeding branch.2.Different date of cutting affected the rooting rate markedly. Withn suitable period, the rooting rate was higher when cutting taken on later date,and the exogenous phytohormone lowered its’ effect. 3.The studies showed several factors, ie. water content, soluble protein, soluble sugar, and polyphenol oxidase activity as well affected the rooting rate. According to coefficient analysis, the rooting rate had negative correlation with bound water, and positive with free water. The water content dropped until 9 days after cutting. soluble protein, soluble sugar, and polyphenol oxidase activity improved slowly. The rate of IAA/ABA and IAA/ZR peaked in 4 to 6 days in cuttings, which means that the content of IAA increased quickly at initial period was very important for cell growth and root primordium formation. 4.The structural investigation of cutting showed that: (1) There was no incubate root primordium in cuttings of Malus prunifolia Borkh, The induced root primordium of developed into the adventitious root, and orginated form the division and differentiation of the vascular cambium cell touching with the pith ray. After 15 days, adventitious root primordium developed into young adventitious roots. (2) In the cutting stem of Malus halliana Roehne, The callus generated after 10 day of cutting and not only form much sclerenchyma, but also form adventitous root primordium, and 40 days more later, adventitious root primordium developed into young adventitious roots.5. During the cutting process, for Malus halliana Roehne, the soluble proteins in cut of 46KD, 39KD, 36KD, 28 KD, 26KD, 24KD are related to the rooting. The soluble proteins in 39KD, 26KD, 24KD control the callus growing and adventitious roots differentiating, consequently, they disappeared as the cutting began to generate adventitious roots. The soluble proteins in 28KD and 31KD appeared during the callus and root developing. It seems that the soluble protein in 46KD in cutting was the possible obstacle to the callus differentiation of the cuttings rooting.For Malus prunifolia, 65KD, 52KD, 43KD, 39KD, 31KD, 28KD, 26KD, 22KD are related to its rooting. The soluble proteins in 65KD, 39KD, 26KD, 22KD control adventitious roots primordium generating and adventitious roots differentiating, and they disappeared as the cutting began to generate adventitious roots.更多还原