食品科学 ›› 2021, Vol. 42 ›› Issue (14): 94-102.doi: 10.7506/spkx1002-6630-20200228-313

• 生物工程 • 上一篇    下一篇

链格孢侵染采后甜瓜病程相关蛋白基因表达及酶活性变化规律

王瑾,白羽嘉,冯作山,朱婉彤,曾禹睿慷,蒋莉莹   

  1. (1.新疆农业大学食品科学与药学学院,新疆果品采后科学与技术重点实验室,新疆 乌鲁木齐 830052;2.新疆农业大学作物学博士后科研流动站,新疆 乌鲁木齐 830052)
  • 发布日期:2021-07-27
  • 基金资助:
    新疆维吾尔自治区自然科学基金青年科学基金项目(2016D01B020)

Gene Expression and Activities of Pathogenesis-Related Enzymes in Postharvest Muskmelons Infected with Alternaria alternata

WANG Jin, BAI Yujia, FENG Zuoshan, ZHU Wantong, ZENG Yuruikang, JIANG Liying   

  1. (1. Xinjiang Key Laboratory for Postharvest Science and Technology of Fruits, College of Food Science and Pharmacy, Xinjiang Agricultural University, ürümqi 830052, China;2. Postdoctoral Mobile Station of Crop Science, Xinjiang Agricultural University, ürümqi 830052, China)
  • Published:2021-07-27

摘要: 以伽师瓜和86-1甜瓜为试材,损伤接种链格孢菌,比较两种甜瓜果皮与果肉抗病性差异。甜瓜果实接种链格孢,(7±1)℃、相对湿度85%~90%冷库贮藏,测定病斑直径,观察病斑组织显微结构,实时聚合酶链式反应比较CmGLU、CmCHT1、CmCHT2相对表达量,测定几丁质酶(chitinase,CHT)和β-1,3-葡聚糖酶(β-1,3-glucanase,GLU)活性。结果表明:链格孢侵染两种甜瓜,伽师瓜病斑直径小于86-1甜瓜,果皮小于果肉,第30天时,86-1甜瓜果皮和果肉病斑直径是伽师瓜的1.28 倍和1.29 倍;观察病斑组织显微结构,发现伽师瓜和86-1甜瓜果皮通过细胞壁增厚抵御链格孢侵染,果肉组织细胞壁抵抗能力较弱,侵染末期果皮与果肉组织细胞壁分层、皱缩,细胞内布满菌丝体和分生孢子,部分果肉细胞解体,无法辨认细胞结构;伽师瓜CHT和GLU基因始终高于86-1甜瓜,果皮高于果肉,CmCHT1相对表达量在侵染初期至侵染中期较高,CmCHT2相对表达量在侵染中期较高,表明病原菌在侵染初期至侵染中期诱导了CHT基因的表达;CmGLU相对表达量在侵染中期至侵染末期较高,表明病原菌在侵染中期至侵染末期诱导了GLU基因的表达;链格孢侵入伽师瓜和86-1甜瓜后,果皮和果肉组织CHT和GLU活性随贮藏时间延长呈升高-降低的变化趋势,伽师瓜高于86-1甜瓜,果皮高于果肉。表明伽师瓜更强的抵御能力与CHT、GLU基因与酶活性密切相关。

关键词: 甜瓜;链格孢;几丁质酶;β-1,3-葡聚糖酶;基因表达;酶活性

Abstract: In this study, Jiashi melons and 86-1 melons were artificially wounded and inoculated with Alternaria alternata to investigate the difference in disease resistance in peel and flesh between the two varieties. The inoculated fruits were stored at (7 ± 1) ℃ and relative humidity (RH) of 85%–90% for up to 30 days. During the storage period, lesion size was measured, lesion microstructure was observed, the expression levels of CmGLU, CmCHT1, CmCHT2 genes were determined by real-time polymerase chain reaction (real-time PCR), and the activities of chitinase (CHT) and β-1,3-glucanase (GLU) were measured. The results revealed that the lesion size in Jiashi melons was smaller than that in 86-1 melons during the whole storage period, and the lesion size in peel was smaller than that in pulp. Moreover, the lesion size in the peel and pulp of 86-1 melons was 1.28 and 1.29 times greater than that in Jiashi melons on day 30. It was also found that the cell structure in the peel of both cultivars was thickened to resist infection by this pathogenic fungus; however, disease resistance of the pulp was weaker than that of the peel. At the late stage of infection, the cell wall in the peel and pulp became stratified and shrunk, and numerous hyphae and spores existed inside the cells. Moreover, some pulp cells were disintegrated so that the cellular structure could not be discerned. The relative expression levels of CHT and GLU genes were higher in Jiashi melons versus 86-1 melons, and in the peel versus the pulp over the entire storage period. The relative expression level of CmCHT1 was higher during the early to middle period, and the relative expression level of CmCHT2 was higher during the middle period, suggesting that infection with A. alternata stimulated the expression of CHT gene. Moreover, the relative expression level of CmGLU was higher during the middle to late period, suggesting that infection with A. alternata stimulated the expression of GLU gene. After infection with A. alternata, the enzyme activities of CHT and GLU in peel and pulp tissues were significantly increased and then decreased with storage time, showing higher values in Jiashi melons relative to 86-1 melons, and in peel relative to pulp. Collectively, these results revealed that Jiashi melons showed stronger disease resistance than 86-1 melons and so did the peel compared with the pulp, which may be closely related with the expression levels and enzyme activities of CHT and GLU.

Key words: muskmelon; Alternaria alternata; chitinase; β-1,3-glucanase; gene expression; enzyme activities

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