纽荷尔脐橙内生真菌参与生理落果的研究

doi:10.16420/j.issn.0513-353x.2022-1000

摘要/Abstract

摘要：

采用ITS高通量测序技术结合真菌分离和培养的方法，研究了纽荷尔脐橙第2次生理落果期间黄化即将脱落的和健康的果实内真菌的多样性，发现即将脱落的果实真菌群落的丰富度、均匀度、物种数显著高于健康果实，并且真菌群落的组成结构也存在显著差异。通过组织分离方法获得了黄化即将脱落的果实中的真菌菌株23株，采用刺伤接种的方法将部分菌株接种于纽荷尔脐橙花后50 d的果实，发现均能引起果实黄化，其中镰刀菌属菌株能够引起幼果脱落酸含量和乙烯释放速率升高。研究结果提示内生真菌群落及引起的内源激素变化可能参与了柑橘第2次生理性落果。

关键词: 柑橘, 落果, 内生真菌, 真菌多样性, Illumina MiSeq, 脱落酸, 乙烯

Abstract:

In order to explore the relation of endophytic fungi and fruit drop，the diversity of fungi in the dropping and healthy fruits of Newhall navel orange during the second fruit drop stage was studied by Illumina MiSeq sequencing technology. It was found that there were significant increases in the richness，evenness of the fungal community in the dropping fruits compared to the healthy fruits，and the composition structure was also different between them. Twenty-three endophytic fungal strains were isolated successfully from the dropping fruits and classified based on ITS sequences. Through artificial inoculation，some endophytic fungal strains could cause yellowing of the young citrus fruits and increase the content of abscisic acid and ethylene release in the fruits. The results suggest that the change of the endophytic fungal communities and the resulting change in the ABA and ETH might be involved in the second drop of citrus fruit.

Key words: citrus, fruit drop, endophytic fungi, fungal community diversity, Illumina MiSeq, abscisic acid, ethylene

崔雪婧, 张家宁, 陈江华, 贾纪春, 林杨, 付艳苹. 纽荷尔脐橙内生真菌参与生理落果的研究[J]. 园艺学报, 2023, 50(12): 2713-2722.

CUI Xuejing, ZHANG Jianing, CHEN Jianghua, JIA Jichun, LIN Yang, FU Yanping. Endophytic Fungi and the Relations to the Fruit Drop of Citrus[J]. Acta Horticulturae Sinica, 2023, 50(12): 2713-2722.

图/表 8

图1 纽荷尔脐橙健康（左）和黄化即将脱落（右）的果实

Fig. 1 The dropping（right） and healthy（left） fruits of Newhall navel orange

表1 纽荷尔脐橙测序数据和质量分析

Table 1 Sequencing data preprocessing statistics and quality analysis of Newhall Navel Orange

果实样品 Sample	原始序列条数 Raw PE	有效序列条数 Clean reads		Q30/%
健康1 Healthy 1	88 653	85 423	231	99.37	96.36
健康2 Healthy 2	95 273	87 349	251	98.93	91.68
健康3 Healthy 3	85 932	82 303	239	99.12	95.78
黄化即将脱落1 Dropping 1	89 633	86 210	240	99.32	96.18
黄化即将脱落2 Dropping 2	93 149	89 388	226	99.24	95.96
黄化即将脱落3 Dropping 3	96 317	92 087	235	99.31	95.61

图2 纽荷尔脐橙测序数据稀释曲线

Fig. 2 ITS sequencing rarefaction curve of Newhall Navel Orange

图3 纽荷尔脐橙健康果实和黄化即将脱落的果实真菌群落的α多样性

Fig. 3 The α-diversity of fungal community in the dropping and healthy fruits of Newhall Navel Orange

图4 纽荷尔脐橙健康果实和黄化即将脱落的果实真菌群落的主成分分析

Fig. 4 PCoA analysis of fungal community in the dropping and healthy fruits of Newhall Navel Orange based on the Weighted Unifrac distance

图5 纽荷尔脐橙健康果实和黄化即将脱落的果实真菌群落属水平物种组成

Fig. 5 The fungal community composition of fungal community in the dropping and healthy fruits of Newhall Navel Orange at the genus level

图6 刺伤接种真菌菌株的纽荷尔脐橙果实

Fig. 6 Influence of fungal strains to Nawhall Navel Orange fruits

图7 纽荷尔脐橙花后50 d的果实接种真菌后11 d脱落酸含量（A）和乙烯释放速率（B） HF：健康果实;DF：黄化果实;NHE-3：链格孢属;NHE-4：毛霉属;NHE-6：节菱孢属;NHE-10：曲霉属;NHE-12：枝孢菌属;NHE-16和NHE-22：镰刀菌属。PDA：对照。单因素ANOVA检验（P < 0.05）。

Fig. 7 Abscisic acid content（A）and ethylene released（B）by citrus fruits 50 d post flower at 11 d post inoculation of fungal strains HF means healthy fruits and DF means dropping fruits. NHE-3：Alternaria sp.;NHE-4: Mucor sp.;NHE-6：Arthrinium sp.;NHE-10：Aspergillus sp.;NHE-12：Cladosporium sp.;NHE-16 and NHE-22：Fusarium sp. PDA：Negative control. One-way ANOVA（P < 0.05）.

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