甘薯病毒病发生关键因素研究

doi:10.3864/j.issn.0578-1752.2021.15.008

摘要/Abstract

摘要：

【目的】明确甘薯种薯携带的病毒种类与苗期病毒病发生率和严重度之间的关系,以及甘薯田烟粉虱发生量和带毒率与种薯带毒率之间的关系,建立甘薯苗期病毒病预测预报方法和种薯质量早期预警技术,为甘薯无病毒种薯生产和病毒病防控提供理论依据。【方法】利用PCR和RT-PCR方法对随机采集的不同来源的甘薯种薯进行病毒检测,检测的病毒种类包括马铃薯Y病毒属（Potyvirus）的甘薯羽状斑驳病毒（sweet potato feathery mottle virus,SPFMV）、甘薯病毒C（sweet potato virus C,SPVC）、甘薯病毒G（sweet potato virus G,SPVG）、甘薯潜隐病毒（sweet potato latent virus,SPLV）和甘薯病毒2（sweet potato virus 2,SPV2）,毛形病毒属（Crinivirus）的甘薯褪绿矮化病毒（sweet potato chlorotic stunt virus,SPCSV）和黄瓜花叶病毒属（Cucumovirus）的黄瓜花叶病毒（cucumber mosaic virus,CMV）以及甘薯双生病毒（sweepoviruses）等我国甘薯上常见的8类主要病毒。然后对检测的种薯进行育苗,出苗后调查薯苗的发病情况,分析种薯携带的病毒种类与苗期病害严重度之间的关系。2018年和2019年分别在河南、宁夏和陕西等地设置试验点,种植背景相同的甘薯品种商薯19原原种苗和脱毒试管苗,在甘薯生长期,采用黄板诱虫法调查各试验点烟粉虱发生量并采集烟粉虱活体,检测烟粉虱SPCSV带毒率。种薯收获后,随机取样对种薯SPCSV带毒率进行检测,分析甘薯田烟粉虱发生量和带毒率对种薯带毒的影响。【结果】在检测的665块甘薯种薯中,有463块种薯携带病毒,育苗后有333块种薯的薯苗表现出叶片黄化、明脉、皱缩和植株矮化等病毒病症状。当种薯携带一种或多种马铃薯Y病毒属病毒时,薯苗病毒病症状主要为0—1级（其中,0级占60.6%;1级占31.8%）;当种薯携带甘薯双生病毒或甘薯双生病毒与马铃薯Y病毒属病毒的组合时, 薯苗病毒病症状主要为0—1级（其中,0级占55.3%;1级占32.9%）;当种薯携带SPCSV时,苗期病毒病症状显著加重,特别是当种薯同时携带SPCSV与马铃薯Y病毒属病毒时,薯苗显症率为100.0%,症状主要为3—9级（其中,3级和5级占49.0%、7级和9级占51.0%）。连续2年田间试验结果表明,甘薯田烟粉虱发生量和带毒率与种薯带毒率密切相关,回归方程为Y=9.628X₁+0.008X₂+6.537,R²=0.914,其中,Y为种薯SPCSV的带毒率,X₁为烟粉虱带毒率,X₂为烟粉虱发生量。【结论】种薯携带SPCSV是苗期病毒病严重发生的关键因素,当种薯同时携带SPCSV与马铃薯Y病毒属病毒时,薯苗病毒病显症率和严重度显著增加。甘薯田烟粉虱发生量和SPCSV带毒率与种薯带毒率密切相关,烟粉虱是影响种薯携带SPCSV的关键因素。

关键词: 甘薯病毒病, 种薯, 烟粉虱, 甘薯褪绿矮化病毒, 预测预报

Abstract:

【Objective】 This study aimed to clarify the relationship between sweet potato virus types in storage roots and virus incidence and severity on root sprouts, and the relationship between numbers and SPCSV-carrying rate of Bemisia tabaci and SPCSV-carrying rate of storage roots, establish the prediction method and early warning technology for viral disease in sweet potato seedlings, so as to provide the theoretical basis for virus-free storage root production, and prevention and control of virus disease in sweet potato.【Method】 PCR and RT-PCR were performed to detect eight major sweet potato viruses including sweet potato feathery mottle virus (SPFMV), sweet potato virus C (SPVC), sweet potato virus G (SPVG), sweet potato latent virus (SPLV) and sweet potato virus 2 (SPV2) in the genus of Potyvirus, sweet potato chlorotic stunt virus (SPCSV) in the genus of Crinivirus, cucumber mosaic virus (CMV) in the genus of Cucumovirus, and sweepoviruses in the genus of Begomovirus in China in storage roots collected from different sources. The PCR-detected roots were planted in plastic pots and allowed to sprout under greenhouse conditions. The types and severity scores of viral symptoms on root sprouts were surveyed and recorded to analyze the relationship between virus species in roots and virus symptom severity on root sprouts. Virus-eliminated sweet potato cultivar Shangshu 19 (S19) plants with the same source were cultivated in the different locations in Henan, Ningxia and Shaanxi in 2018 and 2019. The number of B. tabaci in each test site was calculated by using yellow sticky traps, and was sampled for SPCSV detection during the growth period of sweet potato. The roots selected randomly from each test site were used for SPCSV detection after harvesting to analyze the relationship between numbers and SPCSV-carrying rate of B. tabaci and SPCSV-carrying rate of storage roots.【Result】 Of the 665 storage roots, 463 were infected with one or more of the eight viruses. After root sprouting, visual symptoms, such as leaf chlorosis, vein-clearing, rugosity and stunting were observed on the 333 root sprouts. Virus severity scores on root sprouts ranged from 0 to 1 (60.6% for score of 0 and 31.8% for score of 1) when roots were infected with potyvirus (es). Virus severity scores on root sprouts ranged from 0 to 1 (55.3% for score of 0 and 32.9% for score of 1) when roots were infected with sweepoviruses or co-infected with sweepoviruses and potyvirus (es). Root sprouts were observed with more severe symptoms when roots carried SPCSV, especially the combination of SPCSV and potyvirus (es), which caused visible symptoms on all sprouts with severity scores from 3 to 9 (49.0% for scores of 3 and 5, 51.0% for scores of 7 and 9). Results from two successive years field trial revealed that there was a positive correlation between SPCSV-carrying rate (X₁) and numbers (X₂) of B. tabaci and SPCSV-carrying rate (Y) of storage roots, and the equation of linear regression was Y=9.628X₁+0.008X₂+6.537, R²=0.914.【Conclusion】 Storage root carrying SPCSV is the key factor for serious occurrence of sweet potato viruses at seedling stage. Viral symptom appearance rate and severity scores increase significantly when roots are co-infected with SPCSV and potyvirus (es). There is a positive correlation between SPCSV-carrying rate and numbers of B. tabaci and SPCSV-carrying rate of storage roots, indicating that B. tabaci is the key factor influencing the SPCSV-carrying rate of storage root.

Key words: virus disease of sweet potato, storage root, Bemisia tabaci, sweet potato chlorotic stunt virus (SPCSV), forecast

赵付枚,王爽,田雨婷,乔奇,王永江,张德胜,张振臣. 甘薯病毒病发生关键因素研究[J]. 中国农业科学, 2021, 54(15): 3232-3240.

ZHAO FuMei,WANG Shuang,TIAN YuTing,QIAO Qi,WANG YongJiang,ZHANG DeSheng,ZHANG ZhenChen. An Investigation into Key Factors Influencing the Occurrence of Virus Disease in Sweet Potato[J]. Scientia Agricultura Sinica, 2021, 54(15): 3232-3240.

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目标病毒 Target virus	引物名称 Primer	引物序列 Primer sequence (5′- 3′)	片段大小 Expected size (nt)
Sweepoviruses	SweepoCP-F1	ACTTCGAGACAGCTATCGTGC	645
Sweepoviruses	SweepoCP-R1	ATAYACAGGATTRCTAGCATG	645
SPFMV	SPFMV-F1	TGGGGTTATGATGAACTTCTTC	400
SPFMV	SPFMV-R1	TTCTGGAATGRYTGCGGGTTG	400
SPCSV	SPCSV-F1	TGGGAAGAMGAGAYATGGAG	573
SPCSV	SPCSV-R1	GAGCGTTCCTTTTCATCATC	573
CMV	CMR3-N50	CGCAGGTGGTTAACGGT	697
CMV	CMR3-C30	CCA CACGGTAGAATCAAAT	697
SPVG	SPVG-F1	GCAGGAAAAACAGGAAACAC	792
SPVG	SPVG-R1	ATACCTCGGCATGTATGCCTT	792
SPLV	SPLV-F1	AATGCCAAGGCTACAAGGAGT	553
SPLV	SPLV-R1	CAAGTAGTGTGTGTATGTTCC	553
SPV2	SPV2-F1	TGAACCAGTGCATAGATTGG	770
SPV2	SPV2-R1	TTCACACCTTCATACCATGC	770
SPVC	SPVC-F1	GGACGAACAATGAAAGTTGG	600
SPVC	SPVC-R1	CACTTTCCTGTAATCTGCTG	600

种薯上病毒侵染类型 Virus infection status on storage roots	种薯块数（个） Number of storage roots	各级显症薯苗的比例 Ratio of root sprouts with different virus symptom severities (%)
种薯上病毒侵染类型 Virus infection status on storage roots	种薯块数（个） Number of storage roots	0	1	3-5	7-9
None	202	96.0	3.0	1.0	0
Potyvirus (es)	66	60.6	31.8	7.6	0
Sweepoviruses or sweepoviruses+potyvirus (es)	76	55.3	32.9	11.8	0
SPCSV	142	30.9	29.6	29.6	9.9
SPCSV+sweepoviruses	28	14.3	25.0	46.4	14.3
SPCSV+potyvirus (es)	149	0	0	49.0	51.0
SPCSV+CMV	1	0	0	-	0
CMV	1	0	0	-	0

试验地点 Test site	经纬度 Geographic location	9月28日平均烟粉虱数量（头/黄板） Mean number of B. tabaci	烟粉虱SPCSV带毒率 SPCSV-carrying rate of B. tabaci (%)	种薯SPCSV带毒率 SPCSV-carrying rate of storage roots (%)
原阳试验点 Yuanyang	N35°0′35″,E113°42′31″	276.2	3.0	30.4
延津试验点 Yanjin	N35°15′56″,E114°11′57″	376.8	4.0	47.7
银川试验点 Yinchuan	N38°25′55″,E106°02′28″	80.6	0	14.8

试验点 Test site	经纬度 Geographic location	平均烟粉虱数量（头/黄板） Mean number of B. tabaci			烟粉虱SPCSV带毒率 SPCSV-carrying rate of B. tabaci (%)			烟粉虱平均带毒率 Mean SPCSV- carrying rate of B. tabaci (%)	种薯SPCSV带毒率 SPCSV-carrying rate of storage roots (%)
试验点 Test site	经纬度 Geographic location	8月9日 August 9th	8月29日 August 29th	9月27日 September 27th	8月9日 August 9th	8月29日 August 29th	9月27日 September 27th	烟粉虱平均带毒率 Mean SPCSV- carrying rate of B. tabaci (%)	种薯SPCSV带毒率 SPCSV-carrying rate of storage roots (%)
原阳试验点1 Yuanyang 1	N35°0′35″,E113°42′31″	739.0	5461.2	2398.6	7	0	17	8.0	96.0
原阳试验点2 Yuanyang 2	N35°0′25″,E113°41′28″	380.6	2966.8	4772.2	3	0	5	2.7	74.0
延津试验点1 Yanjin 1	N35°16′27″,E114°12′41″	941.0	2850.0	2236.4	3	2	5	3.3	38.0
延津试验点2 Yanjin 2	N35°15′56″,E114°11′57″	384.0	3286.0	1229.8	2	4	12	6.0	92.0
温县试验点 Wenxian	N35°01′35″,E113°05′17″	1973.0	5098.4	2980.0	1	3	3	2.3	56.0
银川试验点 Yinchuan	N38°25′55″,E106°02′28″	0	537.2	0	0	0	0	0	10.0
榆林试验点 Yulin	N38°29′34″,E109°30′04″	0	0	0	0	0	0	0	0