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[来自光棘豆和变异黄芪的内生真菌尖孢炭疽菌的显微分布及定量检测]

[Microscopic distribution and quantitative detection of endophytic fungus Undifilum oxytropis from Oxytropis glabra DC and Astragalus variabilis].

作者信息

Zhou Qiwu, Yu Longfeng, Lu Hao, Cao Dandan, Zhao Baoyu

出版信息

Wei Sheng Wu Xue Bao. 2014 May 4;54(5):572-81.

Abstract

OBJECTIVE

The characteristics of microscopic distribution and content of Undifilum oxytropis were observed and quantified in different tissues of Oxytropis glabra and Astragalus variabilis from natural grasslands of Inner Mongolia and Ningxia Province.

METHODS

Distribution of fungal endophyte was obtained in all the tissues (stems, leaves, seeds and roots) of O. glabra and A. variabilis though paraffin section and staining method of lactic acid phenol cotton blue; and content of fungal endophyte was determined though Real time-qPCR.

RESULTS

Endophytc fungi were observed mainly within the gap between the palisade tissue and parenchymatous tissue of seed coat in seed, mainly colonized in the superficial cells layer near stoma in the leaves, and in pith of stem mainly plants in the parenchymatous tissue around the edge of the vertical axis of the vascular bundle. There was an obviously difference in concentration of U. oxytropis in the plants collected in different locations. The content of U. oxytropis was highest in all seeds of O. glabra, while it was opposite in stems and leaves of two sampling points. Similarly, the content of U. oxytropis was highest in seeds, it was lowest in roots, and stems and leaves were opposite in A. variabilis from two sampling points. The detection limit was 0.029 pg/ng total DNA by Real time-qPCR.

CONCLUSION

When endophytic fungi infected the tissues of plants, there was selectivity to the tissues and cell type of host, and the colonization and distribution were influenced by habitats in fungal endophytes of locoweeds.

摘要

目的

观察并定量分析内蒙古和宁夏天然草原上光滑棘豆和变异黄芪不同组织中内生丝孢镰刀菌的微观分布特征及含量。

方法

采用石蜡切片法及乳酸酚棉蓝染色法,观察内生真菌在光滑棘豆和变异黄芪所有组织(茎、叶、种子和根)中的分布;采用实时荧光定量PCR法测定内生真菌的含量。

结果

内生真菌主要存在于种子种皮栅栏组织与薄壁组织的间隙中,在叶片中主要定殖于气孔附近的表层细胞,在茎中主要定殖于维管束垂直轴边缘的薄壁组织。不同产地采集的植株中内生丝孢镰刀菌的浓度存在明显差异。光滑棘豆所有种子中内生丝孢镰刀菌的含量最高,而两个采样点的茎和叶中情况则相反。同样,变异黄芪两个采样点的种子中内生丝孢镰刀菌的含量最高,根中最低,茎和叶的情况相反。实时荧光定量PCR的检测限为0.029 pg/ng总DNA。

结论

内生真菌侵染植物组织时,对宿主的组织和细胞类型具有选择性,疯草内生真菌的定殖和分布受生境影响。

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