杉木炭疽菌遗传转化及附着胞发育过程的细胞核行为观察

doi:10.3969/j.issn.1000-2006.201705009

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南京林业大学学报（自然科学版） ›› 2017, Vol. 41 ›› Issue (06): 68-72.doi: 10.3969/j.issn.1000-2006.201705009

杉木炭疽菌遗传转化及附着胞发育过程的细胞核行为观察

黄麟,杨济云,方玉兰,李秋成,梁燕,叶建仁^*

南方现代林业协同创新中心,南京林业大学林学院,江苏南京 210037

出版日期:2017-12-18 发布日期:2017-12-18
基金资助:
基金项目:江苏省高校重大自然科学基金项目(16KJA220002); 江苏高校优势学科建设工程资助项目(PAPD) 第一作者:黄麟(lhung@njfu.edu.cn),副教授。*通信作者:叶建仁(jrye@njfu.edu.cn),教授,博士。

Efficient genetic transformation and nuclear behavior during the process of appressorial development in Colletotrichum gloeosporioides

HUANG Lin, YANG Jiyun, FANG Yulan, LI Qiucheng, LIANG Yan, YE Jianren^*

Co-Innovation Center for the Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, China

Online:2017-12-18 Published:2017-12-18

摘要/Abstract

摘要： 【目的】建立高效的杉木炭疽菌遗传转化体系,并观察附着胞发育过程中的细胞核行为。【方法】通过菌丝体酶解的方法制备原生质体,PEG介导的原生质体转化法将含有伯莱霉素抗性的细胞核表达质粒NL1::GFP转入受体材料杉木炭疽菌SMCG1#C菌株,通过荧光显微镜跟踪观察附着胞发育过程中的孢子、芽管和附着胞等结构中的细胞核行为。【结果】获得了稳定表达NL1::GFP质粒的杉木炭疽菌阳性转化子; 杉木炭疽菌附着胞发育过程伴随着细胞的有丝分裂与细胞核的转移; 附着孢发育成熟后,孢子和芽管菌丝中的细胞核仍然保持完整。【结论】建立了高效的杉木炭疽菌遗传转化体系; 杉木炭疽菌与稻瘟病菌、希金斯炭疽菌的附着胞发育过程的细胞生物学特性显著不同。

Abstract: 【Objective】This study was aimed to establish an efficient genetic transformation system in Colletotrichum gloeosporioides, and to observe the nuclear behavior during the process of appressorial development. 【Method】 Protoplasts were prepared using cell-wall lytic enzymes. Protoplast transformation was mediated by PEG, and the nucleus-specific expression plasmid NL1::GFP with zeocin-resistance gene was transformed into the C. gloeosporioides strain SMCG1#C. During the process of appressorial development, nuclear behavior in the conidia, germ tubes, and appressorium of C. gloeosporioides was observed under a fluorescence microscope. 【Result】Positive C. gloeosporioides transformants expressing NL1::GFP were obtained. Appressorial development in C. gloeosporioides was accompanied by mitosis and cell nuclear transfer. The nuclear in the conidia and germ tubes remained intact without nuclear degration after the appressoria were developed ripely.【Conclusion】 An efficient genetic transformation system was established in C. gloeosporioides. During the process of appressorial development, there were considerable differences in cell-biological characteristics among C. gloeosporioides, Magnaporthe oryzae and C. higginsianum.

中图分类号:

S763
Q945.8

黄麟,杨济云,方玉兰,等. 杉木炭疽菌遗传转化及附着胞发育过程的细胞核行为观察[J]. 南京林业大学学报（自然科学版）, 2017, 41(06): 68-72.

HUANG Lin, YANG Jiyun, FANG Yulan, LI Qiucheng, LIANG Yan, YE Jianren. Efficient genetic transformation and nuclear behavior during the process of appressorial development in Colletotrichum gloeosporioides[J].Journal of Nanjing Forestry University (Natural Science Edition), 2017, 41(06): 68-72.DOI: 10.3969/j.issn.1000-2006.201705009.

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杉木炭疽菌遗传转化及附着胞发育过程的细胞核行为观察

Efficient genetic transformation and nuclear behavior during the process of appressorial development in Colletotrichum gloeosporioides

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