梨轮纹病菌弱毒株携带产黄青霉病毒水平传染力测定

doi:10.16420/j.issn.0513-353x.2022-0683

摘要/Abstract

摘要：

梨轮纹病是由葡萄座腔菌（Botryosphaeria dothidea）引起。前期获得了感染葡萄座腔菌产黄青霉病毒（Botryosphaeria dothidea chrysovirus 1，BdCV1）引起弱毒的梨轮纹病菌菌株LW-C和LW-1，为进一步明确BdCV1介导弱毒菌株是否具有作为候选田间生防资源的潜力，将LW-C和LW-1分别与地理来源不同的梨轮纹病菌（B. dothidea）分离株和危害梨树严重的梨腐烂病菌（Valsa pyri）分离株XJ-28进行对峙培养试验，测定获得携带病毒的衍生菌株生长速率及致病力。研究结果显示轮纹病菌菌株弱毒因子BdCV1均能以一定频率水平传染至不同种类致病菌株中，可引起部分受体菌株生长速率下降，不同程度地抑制其生长。接种梨枝条和果实，原始受体菌株所形成病斑扩展长度约为带毒衍生菌株的2倍至4倍，表明病毒可抑制其受体菌株致病力。以上研究结果揭示了BdCV1可水平传染至不同种类受体菌株中，寄主范围广，具有作为生防因子应用于梨真菌病害中的可行性。

关键词: 梨, 轮纹病菌, 腐烂病菌, 葡萄座腔菌产黄青霉病毒（BdCV1）, 水平传播, 生物防治

Abstract:

Pear ring rot disease，caused by the pathogenic agent of Botryosphaeria dothidea. Botryosphaeria dothidea chrysovirus 1（BdCV1）induced to the hypovirulence of B. dothidea LW-C and LW-1，which will be used as an ideal candidate material for biological control of fruit tree ring rot disease. In order to further clarify whether the BdCV1-mediated hypovirulent strain has potential as a good candidate biocontrol resource，LW-C and LW-1 was confrontly cultured with the B. dothidea strains isolated and collected from different geographical origins and Valsa pyri strain XJ-28，which caused serious harm to pear trees，respectively. The obtained derivate strains were detected to be positive for BdCV1，whose growth rates and pathogenicity were further determined and analyzed. The results showed that BdCV1 could be successfully horizontally transmitted to different B. dothidea and V. pyri strains with certain frequencies，which further inhibited the growth rates and induced to hypovirulence of the strains. The obtained results revealed BdCV1 could be successfully transmitted to the recipient strains at a certain frequency，which could cause the growth rate to decrease and inhibit the growth of the recipient strains to different extent. The formed lesion extension lengths of derivate strains with BdCV1 range from about two to four times longer than that of the recipient strains，when they are inoculated with pear branches or fruits. It further demonstrated that BdCV1 can inhibit the virulence of recipient strains. In summary，BdCV1 could be horizontally transmitted to different recipient strains，revealing that it has wide host range. It is predicted that BdCV1 could be used as a potential biocontrol factor in pear fungal disease biocontrol in practice.

Key words: pear, Botryosphaeria dothidea, Valsa pyri, Botryosphaeria dothidea chrysovirus 1 （BdCV1）, horizontal transmission, biological control

高云静, 王天好, 胡旺成, 洪霓, 王国平, 王利平. 梨轮纹病菌弱毒株携带产黄青霉病毒水平传染力测定[J]. 园艺学报, 2023, 50(11): 2477-2488.

GAO Yunjing, WANG Tianhao, HU Wangcheng, HONG Ni, WANG Guoping, WANG Liping. Determination of Horizontal Transmission Ability of BdCV1 Infecting Botryosphaeria dothidea Strain with Hypovirulence[J]. Acta Horticulturae Sinica, 2023, 50(11): 2477-2488.

图/表 10

表1 对峙培养试验所需受体菌株信息

Table 1 The information of recipient strains used for confrontation culture

病原种类 Pathogen species	菌株编号 Strains number	采集地 Collection place	寄主品种 Host species	枝干症状 Branches symptom
梨轮纹病菌 Botryosphaeria dothidea	HB-2	河北 Hebei	雪青 Xueqing	轮纹 Ring rot
	LN128	辽宁大连 Dalian，Liaoning	道拉恩道梨 Daolaendao pear	干腐、溃疡 Dry rot，ulcer
	HB216	湖北武汉 Wuhan，Hubei	砂梨‘丰水’ Pyrus pyrifolia ‘Housi’	轮纹 Ring rot
梨腐烂病菌Valsa pyri	XJ-28	新疆库尔勒 Korla，Xinjiang	库尔勒香梨 Korla pear	树皮腐烂、黄色卷丝状孢子角 Bark decay，yellow filamentous spore horns

图1 菌株置于PDA或燕麦培养基上水平对峙培养示意图黑色圆点：供体菌株LW-C；蓝色圆点：受体菌株；红色星号：后代衍生菌株。

Fig. 1 The schematic diagram of the horizontal transmission of BdCV1 by confrontation culture of donor strains and recipient strains on PDA or OMA Black dots：Donor strain；Blue dots：Recipient strain；Red pentagram：Derivates.

图2 BdCV1在不同轮纹病菌株中水平传染受体菌株生长观察结果 Ⅰ：LW-C和受体菌株分别在燕麦培养基培养6 d；Ⅱ：LW-C与受体菌株在燕麦培养基对峙培养6 d；Ⅲ：衍生菌株在PDA培养基培养6 d。LW-C/HB-2-1、LW-C/HB-2-3等末位数字表示4组重复中的重复1和3。

Fig. 2 The colony morphologies of strains from horizontal transmission of BdCV1 to different Botrytosperia dothidea isolates Ⅰ：Colony growth of LW-C and recipient strains cultured on OMA for six days；Ⅱ：Colony morphologies of LW-C and recipient strains in confrontation culture on OMA for six days；Ⅲ：Colony morphologies of derivates cultured on PDA for six days. The last digits such as LW-C/HB-2-1 and LW-C/HB-2-3 represent replicates one and three in the four sets of replicates.

图3 衍生菌株dsRNA琼脂糖凝胶电泳检测（A）及生长速率测定（B）不同小写字母表示各处理间差异显著（P < 0.05）。下同。

Fig. 3 The dsRNA profiles detection by 1.2% agarose gel electrophoresis（A）and growth rate determination（B）of derived strains Different lowercase letters indicate significant differences among different treatments at 0.05 level. The same below.

图4 受体菌株及衍生菌株接种5 d‘黄冠’梨果实发病情况

Fig. 4 Inoculation of recipient and derivate strains on‘Huangguan’pear fruit for five days induced to lesion symptoms

图5 接种受体菌株及衍生菌株5 d‘黄冠’梨果实的病斑直径

Fig. 5 The lesion diameter of‘Huangguan’pear fruit at five days after inoculation with the recipient and derivate strains

图6 BdCV1在梨腐烂病菌菌株中的水平传染试验 Ⅰ为LW-1和受体菌株XJ-28分别培养6 d和5 d；Ⅱ为对峙培养5 d；Ⅲ为衍生菌株培养3 d。

Fig. 6 Horizontal transmission test results of BdCV1 in Valsa pyri XJ-28 stain Ⅰ：Colony morphologies of LW-1 and recipient strain XJ-28 cultured on PDA for six days and five days，respectively；Ⅱ：Colony morphologies of LW-1 and recipient strain XJ-28 in confrontation culture on PDA for five days；Ⅲ：Colony morphologies of derivates cultured on PDA for three days.

图7 BdCV1在衍生菌株dsRNA电泳（A）和RT-PCR电泳（B）的结果

Fig. 7 Derived strain dsRNA electrophoresis（A）and RT-PCR electrophoresis（B）results of BdCV1 in Valsa pyri XJ-28 stain

图8 XJ-28及3株衍生菌株生长速率

Fig. 8 Growth rate determination of XJ-28 and three derivate strains

图9 受体菌株及衍生菌株接种‘红香酥’梨枝条症状（A）及7 d病斑长度（B） Ⅰ：空白PDA，Ⅱ：受体菌株XJ-28，Ⅲ：衍生菌株。

Fig. 9 Inoculation of recipient strain XJ-28 and derivates on‘Hongxiangsu’pear branches exhibited lesion symptoms（A） and lesion extension lengths in diameter were determined for seven days（B） Ⅰ：Branches disease symptom of PDA；Ⅱ：Branches disease symptom of recipient strain XJ-28；III：Branches disease symptoms of derived strains.

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