劉瑋琦, 張迎迎, 尹榮玲, 楊永生, 劉文敏, 趙文軍

(1. 滿洲里出入境檢驗檢疫局, 滿洲里 021400; 2. 內(nèi)蒙古大學(xué), 呼和浩特 010021; 3. 呼倫貝爾出入境檢驗檢疫局, 呼倫貝爾 021000; 4. 烏蘭察布出入境檢驗檢疫局, 烏蘭察布 012000;5. 內(nèi)蒙古出入境檢驗檢疫局, 呼和浩特 010020; 6. 中國檢驗檢疫科學(xué)研究院, 北京 100176)

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專論與綜述Reviews

斑馬片病研究進展

劉瑋琦1, 張迎迎2, 尹榮玲3, 楊永生4, 劉文敏5, 趙文軍6*

(1. 滿洲里出入境檢驗檢疫局, 滿洲里 021400; 2. 內(nèi)蒙古大學(xué), 呼和浩特 010021; 3. 呼倫貝爾出入境檢驗檢疫局, 呼倫貝爾 021000; 4. 烏蘭察布出入境檢驗檢疫局, 烏蘭察布 012000;5. 內(nèi)蒙古出入境檢驗檢疫局, 呼和浩特 010020; 6. 中國檢驗檢疫科學(xué)研究院, 北京 100176)

斑馬片病是一種可對馬鈴薯等茄科作物造成嚴重危害的病害,其病原為CandidatusLiberibacter solanacearum,主要通過馬鈴薯木虱(Bactericeracockerelli)等介體進行傳播。該病害主要分布于美國、墨西哥、中美洲各國和新西蘭等國家,雖然目前中國沒有發(fā)生的報道,但具備該病害傳入和發(fā)生的條件。本文對斑馬片病的發(fā)生發(fā)展過程,病原基因組學(xué)、分子檢測技術(shù)、防控措施及對我國的風險等進行討論,對于全面了解該病害,防范其傳入我國并在我國流行具有重要意義。

斑馬片病;CandidatusLiberibacter solanacearum; 馬鈴薯木虱; 分子檢測

斑馬片病(zebra chip,ZC)可危害馬鈴薯、番茄、茄子、綠番茄、樹番茄等茄科作物,是馬鈴薯上的一種毀滅性病害,嚴重影響其產(chǎn)量與品質(zhì)[1],其病原為CandidatusLiberibacter solanacearum,簡稱Lso。此外,歐洲多國報道該病可危害胡蘿卜和芹菜[2]。目前斑馬片病已在北美洲(墨西哥、美國),中美洲(危地馬拉、洪都拉斯、尼加拉瓜),大洋洲[3](新西蘭)和歐洲[4](芬蘭、法國、挪威、西班牙、瑞典)發(fā)生并造成嚴重經(jīng)濟損失。

斑馬片病于1994年在墨西哥東北部城市薩爾提略被首次報道,2000年美國德克薩斯州報道了該病的發(fā)生。短短幾年內(nèi),該病害的發(fā)生面積和頻率大大增加[5],相繼在美國多個地區(qū)發(fā)生[6-7]。墨西哥的大多數(shù)馬鈴薯產(chǎn)區(qū)也發(fā)生了斑馬片病[8],嚴重時發(fā)生率達100%,造成巨大經(jīng)濟損失。2008年在新西蘭發(fā)現(xiàn)該病的流行,60%以上的馬鈴薯田被感染,染病薯塊的品質(zhì)和商業(yè)價值嚴重下降[9-10]。中美洲地區(qū)的危地馬拉、洪都拉斯、薩爾瓦多和尼加拉瓜也已報道該病害的發(fā)生[11]。由胡蘿卜木虱(Triozaapicalis)傳播的斑馬片病在歐洲可造成100%的農(nóng)田損失[12]。

1 危害癥狀

斑馬片病主要危害寄主植物的葉片、枝干和果實等部位,在不同寄主甚至同一寄主不同品種上的危害癥狀及危害程度均有差異。癥狀主要有新葉褪綠黃化,葉片變尖變紫,卷曲,嚴重時葉片卷曲成杯狀(圖1 a和b);莖稈節(jié)間縮短,產(chǎn)生腋生芽,節(jié)點腫大,基部產(chǎn)生氣生薯;受侵染的馬鈴薯塊莖出現(xiàn)壞疽斑和黑色條紋,薯片經(jīng)油炸后黑色條紋會更加明顯,很像斑馬身上的條紋,該病害因此得名斑馬片病[13](圖1c～f)。番茄染病后果實畸形,呈草莓狀,子房發(fā)育不均衡,甚至不結(jié)果,導(dǎo)致產(chǎn)量降低。斑馬片病在葉片上的癥狀與一些木虱引起的黃葉病和馬鈴薯紫頂植原體病害癥狀相似。

與引起柑橘黃龍病的CandidatusLiberibacter相比,Lso對熱更敏感,其最適發(fā)病溫度為27～32℃。當溫度低于或等于17℃時,癥狀發(fā)展會顯著減緩但不阻止Lso在馬鈴薯上的發(fā)育。當溫度高于32℃時,用攜帶Lso的馬鈴薯木虱(Bactericeracockerelli)接種馬鈴薯植株,未觀察到斑馬片病的發(fā)病癥狀,也沒在其上檢測到Lso病原[14]。

圖1 受Candidatus Liberibacter solanacearum 侵染的馬鈴薯葉片和塊莖癥狀[3]Fig.1 Symptoms of potato leaves and tubes infected by Candidatus Liberibacter solanacearum[3]

2 病原

雖然1994年就有斑馬片病發(fā)生的報道,但多年后才明確其病原。2009年Liefting等用電鏡在感染斑馬片病的馬鈴薯和番茄植株韌皮部篩管內(nèi)觀察到了大量大小為0.2 μm×4 μm的類細菌。這些類細菌形狀多樣,并存在一個微弱的可染色中間區(qū)域?；蚍治鲲@示該病菌與柑橘黃龍病菌具有較高的相似性,由于寄主多為茄科作物,故將其命名為CandidatusLiberibacter solanacearum[9]。

2011年,Lin等[15]完成了Lso的全基因組測序(表1),通過比較亞洲柑橘黃龍病菌(Ca. L.asiaticus)、美洲柑橘黃龍病菌(Ca. L.americanus)和Lso三者間的基因組發(fā)現(xiàn),在基因組結(jié)構(gòu)和序列相似性上,與亞洲柑橘黃龍病菌相比,美洲柑橘黃龍病菌與Lso的基因組更為相似。

表1 Candidatus Liberibacter solanacearum 基因組基本特征Table 1 General features of the CandidatusLiberibacter solanacearum genome

Munyaneza等[16]認為Lso可能存在5種地理單倍型(haplotypes)。通過對16S rRNA 的SNPs、16S/23S ISR、50S rplJ和rplL 核糖體蛋白基因進行分析表明:單倍型A和B可引起馬鈴薯和其他茄科作物的斑馬片病[17],由馬鈴薯木虱(B.cockerelli)傳播。其他3種單倍型(C、D和E)可引起胡蘿卜斑馬片病[18-19],由胡蘿卜木虱(T.apicalis)和Bactericeratrigonica傳播。A型主要發(fā)生在洪都拉斯和危地馬拉、遍及墨西哥西部到美國西部的亞利桑那州、加利福尼亞州、俄勒岡州、華盛頓以及新西蘭[20]。B型主要在墨西哥東部,向北遍布德克薩斯州和美國中部。上述兩種單倍型在德克薩斯州、堪薩斯州和內(nèi)布拉斯加州地區(qū)混合發(fā)生。C型在芬蘭、瑞典和挪威的胡蘿卜上發(fā)現(xiàn)[17,19],由胡蘿卜木虱傳播。D和E型主要發(fā)生在西班牙,加納利群島和摩洛哥[19-20],由B.trigonica傳播。目前還不清楚這5種單倍型在寄主植物或昆蟲介體上的生物學(xué)差異。

3 病害的傳播

2007年Munyaneza等[1]首次提出馬鈴薯木虱是斑馬片病的傳播介體,其通過取食染病植物攜帶Lso并進行傳播[16]。Lso在馬鈴薯木虱體內(nèi)的潛伏期為2周左右[21-22]。由于番茄植株中Lso濃度明顯高于馬鈴薯中的濃度[22],當木虱取食染病番茄植株時潛伏期會縮短。馬鈴薯木虱成蟲從攜帶Lso并傳播到馬鈴薯上,到引起馬鈴薯發(fā)病的過程只需要2～6 h[23]。此外,Lso也能通過嫁接侵染馬鈴薯,但傳播速度比通過木虱傳播慢得多。馬鈴薯植株被攜帶Lso的馬鈴薯木虱取食后,在塊莖上表現(xiàn)癥狀一般需要3周左右[24]。斑馬片病可造成馬鈴薯塊莖中含糖量顯著減少,而且癥狀會在儲存過程中繼續(xù)發(fā)展。發(fā)病種薯一般不能發(fā)芽,或產(chǎn)生毛發(fā)狀芽(hair sprouts),植株長勢較弱。

胡蘿卜木虱也是斑馬片病的傳播介體[25]?？梢鸷}卜和芹菜的斑馬片病。最近研究表明Lso可隨胡蘿卜種子傳播[26]。

4 分子檢測方法

目前報道用于Lso的分子檢測方法主要包括常規(guī)PCR、巢式PCR、多重PCR和實時熒光PCR等,效果較好且應(yīng)用較多的PCR方法主要有8種(表2)。

5 防控措施

目前所有防控馬鈴薯斑馬片病的措施都是針對馬鈴薯木虱進行的,加強該蟲的早期監(jiān)測和防控對于減少田間木虱的繁殖和斑馬片病的發(fā)生都具有重要作用[32-33]?？刹捎命S色粘板、水盤、捕蟲網(wǎng)等多種方法對田塊及其周圍的木虱進行監(jiān)測。化學(xué)防治是當前防控馬鈴薯木虱的主要方法。盡管有很多種類的殺蟲劑對該昆蟲有效,但其還是很難控制[34],而且傳統(tǒng)的殺蟲劑只能減少木虱的種群數(shù)量,限制斑馬片病的發(fā)展,卻不能有效控制斑馬片病發(fā)生。

6 斑馬片病的傳入風險及在我國的適生性分析

Munyaneza[16]認為馬鈴薯斑馬片病的傳播必須依靠馬鈴薯木虱,因為植株染病后存活期都很短暫,隨塊莖傳播的可能性很小。然而Pitman等[35]的試驗表明,在沒有馬鈴薯木虱存在的情況下,Lso也可以通過種薯薯塊轉(zhuǎn)移到新生的植株葉片,并能夠傳播給新生馬鈴薯塊莖。該病原物除通過昆蟲介體傳播外,還可通過嫁接、染病植株、帶菌種子和馬鈴薯種薯等進行傳播和擴散。王茹琳等[36]根據(jù)該病害目前的分布和適宜環(huán)境條件,分析了Lso在中國的適生性分布,認為該病在我國的高風險區(qū)為重慶、陜西大部、貴州北部、四川東部、河南大部以及山東半島等地區(qū);中風險區(qū)分布較廣,以高風險區(qū)為中心向外擴散。

馬鈴薯種薯和胡蘿卜種子的國際貿(mào)易都是Lso遠距離傳播的可能途徑,國外檢疫部門高度重視,EPPO(歐洲和地中海植物保護組織)已將該病原菌列為A1類植物檢疫性有害生物[4]。綜合考慮斑馬片病侵入我國的風險:(1)適生范圍廣,危害性大;(2)我國幅員遼闊,完全具備該病害發(fā)生發(fā)展的氣候及寄主條件;(3)其主要寄主馬鈴薯已上升為我國第四大主糧作物,種植面積和產(chǎn)量均居世界第一位[37],經(jīng)濟影響大。因此,斑馬片病一旦傳入將對我國馬鈴薯、番茄、胡蘿卜等產(chǎn)業(yè)造成嚴重威脅。另外,由于馬鈴薯木虱和胡蘿卜木虱自身對寄主的危害不大,檢疫重要性未受到足夠重視,在檢疫過程中容易忽視其作為傳播介體的風險,因此,口岸檢疫中應(yīng)加強來自疫區(qū)的植物產(chǎn)品中傳病昆蟲介體的重視。

[1] Munyaneza J E, Crosslin J M, Upton J E.Association ofBactericeracockerelli(Homoptera: Psyllidae) with zebra chip, a new potato disease in southwestern United States and Mexico [J]. Journal of Economic Entomology, 2007, 100(3): 656-663.

[2] Alfaro-Fernándezm A, Cebrián M C, Villaescusa F J, et al. First report of ‘CandidatusLiberibacter solanacearum’ in carrot in mainland Spain [J]. Plant Disease, 2012, 96(4): 582.

[3] Munyaneza J E.Zebra chip disease of potato: biology, epidemiology, and management [J]. American Journal of Potato Research,2012, 89(5): 329-350.

[4] http:∥www.eppo.int/QUARANTINE/Alert_List/bacteria/Liberibacter_psyllaurous.htm

[5] Abad J A, Bandla M, French-Monar R D, et al. First report of the detection of ‘CandidatusLiberibacter’ species in zebra chip disease-infected potato plants in the United States[J]. Plant Disease, 2009, 93(1): 108-109.

[6] Secor G A, Rivera V V, Abad J A, et al. Association of ‘CandidatusLiberibacter solanacearum’ with zebra chip disease of potato established by graft and psyllid transmission, electron microscopy, and PCR [J]. Plant Disease, 2009, 93(6): 574-583.

[7] Crosslin J M, Olsen N, Nolte P.First report of zebra chip disease and “CandidatusLiberibacter solanacearum” on potatoesin Idaho [J]. Plant Disease, 2012, 96(3): 453.

[8] Munyaneza J E, Sengoda V G, Crosslin J M, et al. First report of ‘CandidatusLiberibacter psyllaurous’ in potato tubers with zebra chip disease in Mexico [J]. Plant Disease, 2009, 93(5): 552.

[9] Liefting L W, Sutherland P W, Ward L I, et al. A new ‘CandidatusLiberibacter’ species associated with diseases of solanaceous crops [J]. Plant Disease, 2009, 93(3): 208-214.

[10]Liefting L W, Perez-Egusquiza Z C, Clover G R G, et al. A new ‘CandidatusLiberibacter’ species inSolanumtuberosumin New Zealand [J]. Plant Disease, 2008, 92(10): 1474.

[11]Bextine B, Aguilar E, Rueda A R, et al. First report ofCandidatusLiberibacter solanacearum on tomato in El Salvador[J]. Plant Disease, 2013, 97(9): 1245.

[12]Munyaneza J E, Fisher T W, Sengoda V G, et al. First report of “CandidatusLiberibacter solanacearum” associated with psyllid-affected carrots in Europe [J]. Plant Disease, 2010, 94(5): 639.

[13]田茜, 牟海青, 趙文軍, 等. 一種引起茄科作物病害的病原:CandidatusLiberibacter solanacearum [J]. 植物檢疫, 2012, 26(2): 31-36.

[14]Munyaneza J E, Sengoda V G, Buchman J L, et al. Effects of temperature on ‘CandidatusLiberibacter solanacearum’and zebra chip potato disease symptom development [J]. Plant Disease, 2012, 96(1): 18-23.

[15]Lin Hong, Lou Binghai, Glynn J M, et al. The complete genome sequence of “CandidatusLiberibacter solanacearum”, the bacterium associated with potato zebra chip disease[J]. PLoS ONE, 2011, 6(4): e19135.

[16]Munyaneza J E.Zebra chip disease,CandidatusLiberibacter, and potato psyllid: A global threat to the potato industry [J].American Journal of Potato Research, 2015, 92(2): 230-235.

[17]Nelson W R, Fisher T W, Munyaneza J E.Haplotypes of “CandidatusLiberibacter solanacearum” suggest long-standing separation[J]. European Journal of Plant Pathology, 2011, 130(1): 5-12.

[18]Teresani G R, Bertolini E, Alfaro-Fernanndez A, et al. Association of ‘CandidatusLiberibacter solanacearum’ with a vegetative disorder of celery in Spain and development of real-time PCR method for its detection [J]. Phytopathology, 2014, 104(8): 804-811.

[19]Nelson W R, Sengoda V G, Alfaro-Fernanndez A O, et al. A new haplotype of “CandidatusLiberibacter solanacearum” identified in the Mediterranean region [J]. European Journal of Plant Pathology, 2013, 135(4): 633-639.

[20]Tahzima R, Maes M, Achbani E H, et al. First report of ‘CandidatusLiberibacter solanacearum’ on carrot in Africa[J]. Plant Disease, 2014, 98(10): 1426.

[21]Cooper W R, Sengoda V G, Munyaneza J E.Localization of ‘CandidatusLiberibacter solanacearum’ (Rhizobiales: Rhizobiaceae) inBactericeracockerelli(Hemiptera: Triozidae)[J]. Annals of the Entomological Society of America, 2014, 107(1): 204-210.

[22]Sengoda V G, Buchman J L, Henne D C, et al.CandidatusLiberibacter solanacearum titer over time in the potato psyllid,Bactericeracockerelli(Hemiptera: Triozidae), following acquisition from infected potato and tomato plants [J]. Journal of Economic Entomology, 2013, 106(5): 1964-1972.

[23]Sandanayaka W R M, Moreno A, Tooman L K, et al. Stylet penetration activities linked to the acquisition and inoculation ofCandidatusLiberibacter solanacearum by its vector tomato potato psyllid[J]. Entomologia Experimentalis et Applicata, 2014, 151(2): 170-181.

[24]Buchman J L, Fisher T W, Sengoda V G, et al. Zebra chip progression: from inoculation of potato plants with liberibacter to development of disease symptoms in tubers[J]. American Journal of Potato Research, 2012, 89(2): 159-168.

[25]Munyaneza J E, Fisher T W, Sengoda V G, et al. Association of “CandidatusLiberibacter solanacearum” with the psyllid,Triozaapicalis(Hemiptera: Triozidae) in Europe [J]. Journal of Economic Entomology, 2010, 103(4): 1060-1070.

[26]Bertolini E, Teresani G R, Loiseau M, et al. Transmission of ‘CandidatusLiberibacter solanacearum’ in carrot seeds[J]. Plant Pathology, 2015, 64(2): 276-285.

[27]Wen A, Mallik I, Alvarado V Y, et al. Detection, distribution, and genetic variability of ‘CandidatusLiberibacter’ species associated with zebra chip complex disease of potato in North America[J]. Plant Disease, 2009, 93(11): 1102-1115.

[28]Hansen A K, Trumble J T, Stouthamer R, et al. A new huanglongbing species, ‘CandidatusLiberibacter psyllaurous’ found to infect tomato and potato, is vectored by the psyllidBactericeracockerelli(Sulc)[J]. Applied and Environmental Microbiology, 2008, 74(18): 5862-5865.

[29]Li Wenbin, Abad J A, French-Monar R D, et al. Multiplex real-time PCR for detection, identification and quantification of ‘CandidatusLiberibacter solanacearum’ in potato plants with zebra chip[J]. Journal of Microbiological Methods, 2009, 78(1): 59-65.

[30]Ravindran A, Levy J, Pierson E, et al. Development of primers for improved PCR detection of the potato zebra chip pathogen, ‘CandidatusLiberibacter solanacearum’[J]. Plant Disease, 2011, 95(12): 1542-1546.

[31]Liefting L W, Veerakone S, Ward L I, et al. First report of ‘CandidatusPhytoplasma australiense’ in potato[J]. Plant Disease, 2009, 93(9): 969.

[32]Henne D C, Workneh F, Wen A, et al. Characterization and epidemiological significance of potato plants grown from seed tubers affected by zebra chip disease [J]. Plant Disease, 2010, 94(6): 659-665.

[33]Butler C D, Trumble J T.Spatial dispersion and binomial sequential sampling for the potato psyllid (Hemiptera: Triozidae) on potato [J]. Pest Management Science, 2012, 68(6): 865-869.

[34]Gharalari A H, Nansen C, Lawson D S, et al. Knockdown mortality, repellency, and residual effects of insecticides for control of adultBactericeracockerelli(Hemiptera: Psyllidae)[J]. Journal of Economic Entomology, 2009, 102(3): 1032-1038.

[35]Pitman A R, Drayton G M, Kraberger S J, et al. Tuber transmission of ‘CandidatusLiberibacter solanacearum’ and its association with zebra chip on potato in New Zealand [J]. European Journal of Plant Pathology, 2011, 129(3): 389-398.

[36]王茹琳, 高曉清, 姜淦, 等. 茄科作物新病原CandidatusLiberibacter solanacearum入侵我國的風險分析[J]. 江西農(nóng)業(yè)學(xué)報, 2015, 27(1): 42-46.

[37]羅其友, 劉洋, 高明杰, 等. 中國馬鈴薯產(chǎn)業(yè)現(xiàn)狀與前景[J]. 農(nóng)業(yè)展望, 2015 (3): 35-40.

(責任編輯：田 喆)

Research progress on zebra chip disease

Liu Weiqi1, Zhang Yingying2, Yin Rongling3, Yang Yongsheng4, Liu Wenmin5, Zhao Wenjun6

(1. Manchuria Entry-Exit Inspection and Quarantine Bureau, Manchuria 021400, China; 2. Inner Mongolia University, Hohhot 010021, China; 3. Hulunbuir Entry-Exit Inspection and Quarantine Bureau, Hulunbuir 021008, China; 4. Ulanqab Entry-Exit Inspection and Quarantine Bureau,Ulanqab 012000, China; 5. Inner Mongolia Entry-Exit Inspection and Quarantine Bureau,Hohhot 010020, China; 6. Chinese Academy of Inspection and Quarantine, Beijing 100176, China)

Zebra chip, caused byCandidatusLiberibacter solanacearum, is a kind of serious disease on potatoes and other solanaceous crops by reducing both yield and quality. It is mainly transmitted by potato psyllids (Bactericeracockerelli) and other vectors. The disease is mainly distributed in the United States, Mexico, Central American countries and New Zealand and other countries. Now this disease has not been reported in China, but the environmental conditions of China is suitable for its invasion and occurrence. In this paper, we described the occurrence, pathogen genomics, molecular detection, prevention and control measures. This will be useful for comprehensive understanding the disease, and preventing its spread to our country.

zebra chip;CandidatusLiberibacter solanacearum;Bactericeracockerelli; molecular detection

2015-11-19

2015-12-28

質(zhì)檢總局科技計劃項目(2014IK290)；“十二五”國家科技支撐計劃(2012BAK11B02)

S 435.32

A

10.3969/j.issn.0529-1542.2016.05.005

* 通信作者 E-mail: wenjunzhao@188.com