高级检索+

烟草雌蕊的花粉管生长途径中几种植物激素的免疫金电镜观察

IMMUNOGOLD ELECTRON MICROSCOPIC OBSERVATIONSON DISTRIBUTION OF SEVERAL PHYTOHORMONES IN POLLEN TUBE GROWTH TRACK THROUGH PISTIL IN TOBACCO

  • 摘要: 用免疫金电镜法观察了大叶烟花柱、珠孔及助细胞内玉米素(t-Z)、吲哚乙酸(IAA)、赤霉素(GA7与GA4)和脱落酸((+)ABA)的分布。刚授粉时的花柱引导组织细胞内与胞间基质中t-Z较多、IAA、GA7/4、(+)ABA较少;授粉后24h(此时花粉管已穿过花柱中部),该部位t-Z显著减少,IAA略增加,GA7/4显著增加,而(+)ABA仍较少。授粉后24h,花柱中段薄壁组织细胞内与胞间基质中IAA比刚授粉时显著增加。增加的IAA、GA7/4主要分布在淀粉体、液泡与胞间基质中。穿过花柱中部的花粉管壁上有IAA。刚授粉时珠孔与助细胞内t-Z较多,丝状器内有大量的t-Z;授粉后96h,这些部位的t-Z显著减少。刚授粉时珠孔与助细胞内IAA、GA7/4、(+)ABA较少;授粉后96h,除退化助细胞与宿存助细胞内GA7/4较多外,其它部位的这些激素仍较少。根据这些结果,讨论了雌蕊内这4类植物激素的分布特点和花粉管定向生长的关系

     

    Abstract: Up to now,little is known about distribution and function of phytohormones in the pollen tube growth track through pistils.Distribution of tran-szeatin (t-Z),indele-3-acetic acid (IAA),gibberellins A7and A4 (GA7/4),and(+) abscisic acid((+) ABA)in the style,micropyle and synergids of N icotiana tabacum var.macrophylla were observed with immunogold electron microscopy.Segments from middle part of the styles and ovules at various stages at and after pollination were fixed with 2 % EDC(1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide)and mixed paraformaldehyde and glutaraldehyde for acidic phytohormones (or only with the latter for t-Z),then stightly postfixed in 0.5 % OSO4solution for 30 min.After dehydration,samples were embedded in Epon 812 resin.Following etched in 10 % H2 O2 for 10 min,ultrathin sections were immunostained respectively with rabbitanti t-Z polyclonal antibody (PAb),anti-IAA methyl ester PAb,mouse anti-GA monoclonal antibody(MAb,specifically recognizes methyl esters of GA7and GA4),or anti(+)-ABA methyl ester MAb.Protein Acolloidal gold (Φ10 nm) was used as a marker for rabbit PAbs while sheep antimouse lg Gcolloidal gold(Φ10 nm) for mouse MAb.At pollination plenty of t-Z but a few IAA and GA7/4 were observed in cells and intercellular matrix of the transmitting tissue.One day after pollination,when pollen tubes had penetrated the transmitting tissue,the gold granules of t-Z remarkably decreased,but those of IAA slightly increased; and those of GA7/4 obviously increased in the same region.IAA in cells and intercellular matrix of stylar parenchyma greatly increased from just at pollination to one day after pollination.Increased IAA and GA7/4 were mainly localized in amyloplasts,vacuoles and intercellular matrix.IAA appeared in the walls of pollen tubes passing the transmitting tissue.High level of t-Z in themicropyle and the synergids(particularly abundant t-Z in the filifrom apparatus) greatly decreased from just atpollination to 4 days after pollination.GA7/4 were less in the synergids at pollination butincreased in both the degenerate and persistent synergids 4 days after pollination.GA7/4 and IAA in the micropyle or IAA in the synergids maintained at lower levels-4 days after pollination.(+) ABA in the pollen tube growth track was rarely observed during the whole course.According to the results,the relationship of the distribution mode of these four phytohormones in tobacco pistil with pollen tube guidance is discussed.

     

/

返回文章
返回