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      新疆紅棗棗果主要致病菌交鏈格孢菌產生毒素種類及黑斑病病果毒素含量測定

      2017-08-30 17:35:02何麗郭開發(fā)艾尼古麗依明趙思峰
      新疆農業(yè)科學 2017年6期
      關鍵詞:孢霉鏈格棗果

      何麗,郭開發(fā),艾尼古麗·依明,趙思峰

      (新疆綠洲農業(yè)病蟲害治理與植保資源利用自治區(qū)高校重點實驗室/石河子大學農學院,新疆石河子 832003)

      新疆紅棗棗果主要致病菌交鏈格孢菌產生毒素種類及黑斑病病果毒素含量測定

      何麗,郭開發(fā),艾尼古麗·依明,趙思峰

      (新疆綠洲農業(yè)病蟲害治理與植保資源利用自治區(qū)高校重點實驗室/石河子大學農學院,新疆石河子 832003)

      【目的】研究新疆紅棗縮果病和棗果黑斑病兩種病害的主要致病菌交鏈格孢菌(Alternariaalternate)產生的主要毒素種類,以及栆果黑斑病病果中的毒素種類和含量,為紅棗加工和食用的安全性評價提供依據。【方法】采用超高效液相色譜-串聯(lián)質譜法,測定10株鏈格孢菌產生的毒素種類及含量,并對棗果黑斑病不同嚴重度發(fā)病棗果和人工接種果實中毒素種類及含量進行測定?!窘Y果】10株交鏈格孢供試菌株可產生4種毒素,分別為鏈格孢酚(alternariol,AOH)、交鏈格孢酚單甲醚(alternariol monomethyl ether,AME)、交鏈孢烯(altenuene,ALT)和細交鏈格孢菌酮酸(tenuazonic acid,TeA)。TeA、ALT、AME和AOH在不同病級發(fā)病棗果和人工培養(yǎng)條件下均有檢出,其中TeA檢出含量最高,范圍為3.1×103~5.5×103mg/kg;AME、AOH和ALT的含量范圍分別為7.2×102~6.4×102mg/kg,1.2~3.8×102mg/kg和0.09~5.08 mg/kg。人工接種鏈格孢菌后,無傷、有傷棗果內均檢測到大量TeA。不止在發(fā)病棗果中檢測到了毒素,無傷接種未發(fā)病棗果亦發(fā)現(xiàn)大量毒素,且未接種健康棗果中亦有少量鏈格孢霉毒素?!窘Y論】新疆紅棗棗縮果病和棗果黑斑病病原菌兩種病害的主要致病菌交鏈格孢菌(Alternariaalternate)均可產生鏈格孢霉毒素,且產毒量高,危害嚴重,影響了新疆紅棗的產量及商品價值,患病棗果無法安全食用。在紅棗鮮食加工和風險評估中應引起關注和重視,并在紅棗種植和貯存期間減少病原菌侵染,以避免更多的污染。

      紅棗;棗果縮果??;棗果黑斑??;鏈格孢霉;毒素

      0 引 言

      【研究意義】鏈格孢屬真菌已報道約500種,其廣泛存在于水果、蔬菜和田間作物上導致多種農作物病害,同時該屬真菌還可以產生多種真菌毒素,主要種類有鏈格孢酚(AOH)、交鏈格孢酚單甲醚(AME)、交鏈孢烯(ALT)和細交鏈格孢菌酮酸(TeA)等[1-3],這些毒素具有誘變性、致癌性、基因毒性等多種毒性作用[4]。【前人研究進展】目前已在蘋果、柑橘、番茄、土豆、小麥、葵花籽等農產品中檢測到AME、AOH等鏈格孢毒素[5-8]。Dong等[9]從林縣的谷物上測出多種鏈格孢霉毒素,并認為AOH和AME兩種毒素很有可能與中國河南林縣的食道癌發(fā)生密切相關。AOH能引起食道上皮細胞增殖,并能引起胚胎食道鱗狀細胞癌變,用AOH處理老鼠皮下,可誘導鱗狀上皮細胞癌[10]。AME作為直接誘變劑,可使姊妹染色體交換率上升,誘導NIH 3T3細胞轉化,轉化后的細胞易癌變[11];AME也可選擇性作用于特定的基因組或DNA序列引發(fā)誘變[12]。TeA在鏈格孢霉毒素中毒性最強,具多種生物特性,如抗病毒[13]、抗腫瘤、抑菌[14]、細胞毒性、植物毒性[15],抑制蛋白質的合成[16]等。TeA對哺乳動物(小鼠、大鼠等[17])具有急性毒性,其鈉鹽、鎂鹽與一種發(fā)生在非洲的人類出血性疾病-奧尼賴病(Onyalai)密切相關[18]。目前歐盟已就食品與飼料中鏈格孢霉毒素對人畜的健康進行風險發(fā)布,其中TeA已列入美國食品藥物管理局(FDD)有毒化學物質登記冊?!颈狙芯壳腥朦c】新疆棗縮果病和棗果黑斑病的主要病原菌均為交鏈格孢菌(Alternariaalternate)[19,20],而其是否產生鏈格孢菌毒素以及毒素的種類和量目前尚未有明確報道?!緮M解決的關鍵問題】研究采用超高效液相色譜-串聯(lián)質譜法,對分離自新疆棗縮果病和棗果黑斑病的交鏈格孢菌毒素種類及含量進行測定和分析,并測定及分析棗果黑斑病不同發(fā)病病級的果實和人工接種交鏈格孢菌后紅棗中毒素種類和含量,為新疆紅棗果實的安全食用風險評估提供依據。

      1 材料與方法

      1.1 材 料

      1.1.1 供試菌株

      鏈格孢菌代表菌株(Alternariaalternate)10株,分離自新疆紅棗主產區(qū)的棗縮果病和棗果黑斑病病樣上,經形態(tài)學和分子生物學方法鑒定為交鏈格孢菌(A.alternate),菌種保藏于石河子大學新疆綠洲農業(yè)病蟲害治理與植保資源利用自治區(qū)高校重點實驗室。

      1.1.2 發(fā)病棗果樣品

      采自新疆阿拉爾市周邊駿棗發(fā)病果實,按發(fā)病面積進行分級:0級,果實表面沒有病斑;1級,病斑面積<25%;2級,病斑面積25%~50%;3級,病斑面積50%~75%;4級,病斑面積>75%[21]。

      1.1.3 接種用棗果

      采自阿拉爾市周邊駿棗膨大期果實,快遞至石河子市實驗室內供接種用。

      1.1.4 培養(yǎng)基

      DRYES培養(yǎng)基[22]:酵母膏 20 g、蔗糖150 g、虎紅0.5 g、氯霉素0.1 g(0.2%溶于乙醇)、水1 L,pH 6.8。

      PDA培養(yǎng)基[23]:馬鈴薯(去皮) 200 g、葡萄糖 20 g、瓊脂粉 15 g、水1 L,pH 6.8。

      交鏈格孢酚單甲醚(AME,純度>98%)和鏈格孢酚(AOH,純度>98%)標準品購買于美國Sigma公司;細交鏈格孢菌酮酸(TeA,純度>98%)和交鏈孢烯(ALT,純度>98%)標準品購買于北京萊耀生物公司;其余乙酸乙酯、乙二胺-N-丙基硅烷(PSA)、乙腈等試劑購買于石河子市試劑公司。

      Xevo TQ/MS超高效液相色譜串聯(lián)質譜儀(美國Waters公司),配有Acquity UPLC EH C18液相色譜柱(2.1 mm×150 mm,1.7 μm)。

      1.2 方 法

      1.2.1 超高效液相色譜-串聯(lián)質譜條件

      色譜柱:Acquity UPLC BEH C18液相色譜柱(1.7 μm,50 mm×2.1 mm);流動相:乙腈-水(0.1%甲酸);離子化模式:電噴霧離子源,正離子模式(ESI+);質譜掃描方式:多反應監(jiān)測(MRM);毛細管電壓3.5 kV,錐孔電壓40 kV,離子源溫度150℃,霧化溫度500℃,脫溶劑氣流量1 000 L/h,錐孔氣流速50 L/h。列出4種鏈格孢霉毒素的監(jiān)測離子、錐孔電壓和碰撞電壓等質譜參數。表1

      表1 4種鏈格孢霉毒素的串聯(lián)質譜測定參數
      Table 1 MS/MS parameters for the 4 Alternaria mycotoxins

      成分Compound電離模式IonizationMode母離子Precursor(m/z)子離子Daughter(m/z)保留時間Dwell(s)毛細管電壓Cone(kV)錐孔電壓Collision(kV)AOHESI+259.2213.2185.2*1.093.53.54040AMEESI+273.2199.1128.2*2.33.53.54040TeAESI+198.2153.2125.2*1.123.53.54040ALTESI+293.2257.2257.3*0.823.53.54040

      注:*定量離子

      Note:*Quantitative ion

      1.2.2 10株交鏈格孢菌毒素種類及含量測定

      將10株交鏈格孢菌菌株在PDA培養(yǎng)基上活化后,用打孔器打5 mm菌餅,接種到DRYES培養(yǎng)基上,三次重復,以空白為對照,28℃恒溫、12 h晝夜交替培養(yǎng)14 d后,分別從三次重復中取1 g菌餅,用液氮研磨充分,于2 mL離心管中。離心管內加入1 mL乙酸乙酯(含1%甲酸),震蕩1 h,在4 000 r/min下離心5 min,將提取液轉入一個新2 mL離心管內,以氮吹器蒸發(fā)干燥,將凍干粉末溶解到400 μL甲醇,取上清液過0.45 μm有機濾膜,濾液經UPLC-MS/MS分析。

      1.2.3 不同病級發(fā)病棗果樣品處理

      將采集的不同發(fā)病級數紅棗栆果樣品用清水洗凈晾干,用75%酒精表面消毒,再用無菌水將果實沖洗干凈,最后用滅菌濾紙吸干果實上殘留的水分,分別稱取健康棗果和發(fā)病果實病斑附近果肉2 g(精確至0.01 g),于50 mL尖底具塞離心管中,加入10 mL 1%甲酸乙腈溶液,7.5 mL無菌水,震蕩4 min;再加入4 g無水MgSO4和1 g NaCl,劇烈震蕩2 min 后,在4 000 r/min下離心5 min;取上清液轉入15 mL離心管,加入0.2 g PSA和0.6 g無水MgSO4,劇烈震蕩2 min后,在4 000 r/min下離心5 min;取上清液以氮吹器蒸發(fā)干燥,將凍干粉末溶解到400 μL甲醇,過0.45 μm有機濾膜,濾液經UPLC-MS/MS分析。10株棗縮果病和棗果黑斑病代表性的致病菌株在DRYES培養(yǎng)基上培養(yǎng)14 d后,提取粗提取物,測量4種鏈格孢霉毒素。

      1.2.4 人工接種鏈格孢后發(fā)病棗果樣品處理

      將活化的10株采用離體菌塊貼接法[20],對健康棗果進行無傷、有傷接種,將活化后的供試菌株,用打孔器打5 mm菌餅,將采集的健康無傷的紅棗果實用清水洗凈晾干,用75%酒精表面消毒,再用無菌水將果實沖洗干凈,最后用滅菌濾紙吸干果實上殘留的水分,用滅菌牙簽刺孔,在超凈工作臺中將菌餅菌面朝下接種到紅棗果實上,以DRYES培養(yǎng)基塊為對照,然后放入鋪有濕潤濾紙的方形發(fā)芽盒內(12 cm×12 cm×1.5 cm)中,每個代表菌株接種3個果實,28℃條件下12 h光照、12 h黑暗交替培養(yǎng),5 d后稱取病斑附近果肉2 g,按1.2.3方法處理后測定4種鏈格孢霉毒素種類及含量。

      試驗分別準確稱取健康果肉、DRYES培養(yǎng)基,在20、100、500和1 000 ppb 4個水平下加標,每個水平重復測定6次,按1.3.2樣品處理方法處理并測定,以進樣質量濃度X(ng/mL)為橫坐標、峰面積Y為縱坐標繪制標準曲線。

      2 結果與分析

      2.1 4種鏈格孢霉毒素總離子流

      在乙腈-水(0.1%甲酸)流動相體系中質量濃度為100 ng/mL的混標溶液(AOH、AME、ALT和TeA混合物)在3 min內達到有效分離且峰型良好。保留時間為0.81和2.3 mim的物質分別為ALT和AME,1.09和1.12 min分別為AOH和TeA。4種鏈格孢霉毒素的總離子流中有3個顯示峰,AOH和TeA聚集在一個峰上,但試驗采用超高效液相色譜質譜連用技術,結合濃度為100 ng/mL標準溶液中4種鏈格孢霉毒素(AOH、AME、ALT和TeA)的MRM定量離子對質譜圖,以離子比為參照確定混合物中組分,并不影響試驗結果,試驗可行。圖1,圖2

      圖1 4種鏈格孢霉毒素的混合標準溶液(100 ng/mL)總離子流
      Fig.1 Total ion current of 4 mycotoxins mixed standard solution(100 ng/mL)

      2.2 方法驗證

      4種鏈格孢霉毒素在2~100 ng/mL范圍內均有良好的線性關系,R2>0.99,平均回收率在70%~95%,相對標準偏差均小于8.9%在S/N=3時,ALT、AME、AOH和TeA的檢出限在5~20 ppb。研究表明,該方法對紅棗樣品的不同含量的4種鏈格孢霉毒素的測定均具有較高的回收率和精密度,滿足檢測要求。表2,表3

      2.3 10株鏈格孢菌菌株中毒素種類及含量

      研究表明,10株菌均可產生TeA,在4種鏈格孢霉毒素中TeA含量最高,其中菌株21-HB含量最高,可達5.39×103mg/kg,hm-16產毒最低,含量為35.9 mg/kg;菌株92-3-1和hm-24產生的毒素種類最多,4種鏈格孢霉毒素均可產生;菌株30-HB-2和36-3-3產生毒素種類最少,僅產生TeA;菌株hm-16、hm-31和hm-35可產生AOH、AME、ALT,產毒范圍分別為5.52~3.82×102mg/kg,1.96~7.16×102mg/kg,0.22~2.53 mg/kg。表3

      注:A.ALT;B.AOH;C.TeA;D.AME

      Notes:A.ALT; B.AOH; C.TeA; D.AME

      圖2 4種鏈格孢霉毒素混合標準溶液在ESI+模式下MRM色譜


      Fig.2 Mutiple reaction monitoring (MRM) chromatograms of mixed solution

      of 4 mycotoxins standards in ESI+

      表2 4種鏈格孢霉毒素的線性范圍、線性方程、R2、檢出限(LOD)


      Table 2 Linear equations, correlation coefficients, linear ranges, detections limi, recoveries and precisions for the 4Alternariamycotoxins

      基質成分Components線性方程R2線性范圍(ng/mL)檢出限(LOD)回收率Recovery(%)RSD(%)CKAOHAMETeAALTY=402.347X+2014.14Y=240.212X+1292.26Y=889.202X+12622.6Y=125.01X+227.1710.98510.99130.97630.99262~1001020510847688864.54.73.28.9果肉AOHAMETeAALTY=253.954X+2974.11Y=204.011X+1456.73Y=720.905X+4860.95Y=171.612X+7196.060.98510.99490.99060.98912~1001020510707295844.46.55.97.1培養(yǎng)基AOHAMETeAALTY=253.371X+1116.79Y=191.023X+1620.98Y=728.674X+20711Y=192.937X+302.4590.99790.9980.94840.92942~1001020510736987795.25.96.16.3

      表3 DRYES培養(yǎng)基中10株鏈格孢菌產生毒素種類及含量
      Table 3 The species and content of 10Alternariastrains produce mycotoxins in DRYES medium (mg/kg)

      菌號成分CK34T11L30-HB-2hm-35hm-1692-3-1hm-2421-HB36-3-3hm-60hm-31AOH40.13381.5811.895.5212.8955.70AME12.0264.9836.561.96716.10375.42TeA1.751261.5744.06429.935.882396.634647.965389.21537.31724.404943.01ALT2.530.220.52

      2.4 不同病級發(fā)病棗果中4種鏈格孢霉毒素含量

      研究表明,不同發(fā)病程度發(fā)病棗果中4種鏈格孢霉毒素均有檢出。隨著棗果發(fā)病程度增加,鏈格孢霉毒素含量逐漸升高,3級時毒素含量最高;4種毒素中TeA含量最高,最高可達3.1×103mg/kg,AME、AOH、ALT分別可達6.4×102mg/kg,94.5 mg/kg,16.5 mg/kg。且0級果實中也檢測到了4種毒素,但檢出量較低,可能是運輸儲藏過程中健康棗果受到病果污染所致。表4

      表4 不同病級發(fā)病棗果中4種鏈格孢霉毒素含量
      Table 4 The content of 4Alternariamycotoxins in effected jujube fruit (mg/kg)

      病級TeAAOHAMEALT030.910.040.620.031126.970.401.790.182524.9977.22638.643.3333102.2794.51995.3816.484186.463.092.131.68

      2.5 人工接種棗果后4種鏈格孢霉毒素含量

      研究表明,10株菌人工接種健康棗果后,無傷、有傷棗果內均產生大量鏈格孢霉毒素,其中TeA含量最高,4種毒素含量范圍分別為4.6~2.1×103mg/kg,4.7~1.4×103mg/kg,1.2~122 mg/kg,0.09~5.08 mg/kg。用菌株hm-24接種的產毒量最高,TeA為2 102.90 mg/kg,AME為1 390.86 mg/kg;hm-16產毒量最低,TeA僅為5.40 mg/kg;34T11L、hm-35、hm-24、hm-604個菌株接種后可產生4種鏈格孢霉毒素;30-HB-2、92-3-1、21-HB、和36-3-3接種后在紅棗果肉中僅檢測出了TeA。且CK果肉中也檢測到TeA,可能是樣品在運輸儲藏的過程中健康棗果受到病果污染。表5

      表5 棗果接種后4種鏈格孢霉毒素含量測定
      Fig.5 The content of four Alternaria mycotoxins after inoculated withA.alternateon Jujube fruit (mg/kg)

      成分菌號TeAAOHAMEALTCK-+1.202.8034T11L-+4.6046.407.1032.000.391.2230-HB-2-+16.1041.50hm-35-+54.00429.904.7030.100.09hm-16-+4.505.401.20122.20249.1892-3-1-+5.6070.40hm-24-+17.002102.900.4999.101390.860.685.0821-HB-+6.60151.7036-3-3-+6.2071.10hm-60-+11.8012.207.2922.021.19hm-31-+10.601455.402.90

      注:空白表示未檢測到毒素

      Note:Blank said undetectedAlternaria Mycotoxins

      3 討 論

      鏈格孢菌種類繁多,適應性強,寄主范圍廣,95%以上的種能兼性寄生于植物上,可引起多種植物尤其是糧食作物、蔬菜、水果等農作物和經濟作物病害。A.tenuissima、A.alternata、Alternariasp、A.triticina等可侵染引起小麥葉斑病等[24],A.arborescens、A.tenuissima、A.mali、A.alternata/A.longipes等[25]多種鏈格孢菌可引起蘋果葉斑病和果斑病。A.tenuissima侵染引起的小麥黑胚病(Black point of wheat)是一種世界性籽粒病害,在中國、美國、墨西哥、加拿大、印度、英國、前蘇聯(lián)、秘魯等國均有發(fā)生[26,27]。A.alternata侵染引起的梨黑斑病(裂果病)是梨的三大病害之一,是梨樹上廣泛發(fā)生的世界性病害,尤其在亞洲的日本、韓國和中國發(fā)病十分嚴重[28]。梨黑斑病主要侵染葉片,造成大量落葉;有時也可危害果實和新梢,能造成較大的經濟損失,發(fā)生嚴重時可使梨采后損失達50%以上[29]。2004 年我國鴨梨曾因黑斑病被停止出口美國和加拿大[30]。史文景[31]、程月萌等[32]測定了蘋果、桃和梨中鏈格孢霉毒素,檢測結果表明,蘋果中檢測到AOH和AME,含量分別為49和30 μg/kg。Birgitte等[33]運用超高液相色譜質譜串聯(lián)技術測定了小麥、番茄、核桃、藍莓中的鏈格孢霉毒素,結果表明均存在AOH、AME、ALXII、ALT、TeA、TEN等毒素。Peter 等[34]測定了谷物中鏈格孢霉毒素,檢測結果表明83份樣品中均檢測到AOH、AME,含量分別為0.34和0.13 ng/g。Miller等[17]對小鼠分別經靜脈注射、腹腔、皮下、口腔單次給TeA鈉鹽的LD50分別為(125±10)、(150±10)、(145±20)、(225±25) mg/kg;AOH和AME對實驗動物的急性毒性較弱,小鼠的LD50>100 mg/kg。鏈格孢霉毒素在全球多種農產品中均已檢測到,對人類和動物健康的潛在危害已不容忽視。

      紅棗種植目前已成為新疆近年來發(fā)展最快、效益最突出和惠民成效最為顯著的一項林果產業(yè)[35,36],然而棗果黑斑病在很多地區(qū)均有發(fā)生。研究以新疆紅棗縮果病和棗果黑斑病病果分離的10株交鏈格孢菌以及不同發(fā)病級數的棗果作為研究對象,10株供試菌株均能產生鏈格孢菌毒素,不同菌株產生的毒素種類以及含量有差異,其中從哈密分離到的hm-24菌株可以產生4種毒素,且產生毒素的量也最高,用其接種健康棗果后也可以在果肉中檢測到4種毒素。不同病級棗果中4種鏈格孢霉毒素均有檢出,TeA、AME、AOH、ALT含量分別為30.9~3.1×103mg/kg,0.6~6.4×102mg/kg,0.3~94.5 mg/kg,0.2~16.5 mg/kg。這與Magnani等[30]蔣黎艷等[31]檢測到鏈格孢霉毒素中TeA含量最高而相一致。其中在健康棗果上也檢測到了微量的TeA毒素,可能是運輸過程中污染所致,也有可能健康果實表面也有鏈格孢菌的污染產生的毒素。

      4 結 論

      鏈格孢菌毒素分布面廣,對人以及哺乳動物均有誘變性、致癌性、基因毒性等多種毒性作用,研究采用超高效液相色譜-串聯(lián)質譜法,明確了引起棗縮果病和棗果黑斑病的主要致病菌交鏈格孢菌可以產生TeA、AME、AOH和ALT 4種毒素,毒素含量分別為1.2~2 102.9、22.02~1 390.86、1.2~122.2和0.39~5.08 mg/kg,其中TeA毒素的產生量最高,最高可達2 102.9 mg/kg。用其接種健康棗果5 d后,可在接種部位檢測到大量鏈格孢菌毒素;同時對田間自然條件下采集的不同發(fā)病級數的棗果黑斑病果實進行檢測后也能檢測到4種鏈格孢菌毒素,在健康未發(fā)病的果實上也檢測到了少量鏈格孢菌毒素。因此,安全、有效、及時控制棗果黑斑病已成為新疆紅棗種植過程中的一個影響紅棗食用安全性的關鍵問題,采收時應當將健康果實與病果分開,防止健康果實被病果污染。

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      YANG Xiao-ping, HU Hong-ju, WANG You-ping, et al. (2009). Biological characteristics and pathogencity of pear black spot byAlternariaalternate(Fr.) Keissl [J].JournalofHuazhongAgriculturalUniversity, (12):680-684. (in Chinese)

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      XIE Li, ZHANG Duo, ZHANG Li-ping, et al. (2008). Isolation and identification of an antagonisticStreptomycesstrain againstAlternariaalternate. [J].JournalofHebeiNormalUniversity(NaturalScienceEd.) , 32(4): 526-529. (in Chinese)

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      ZHANG Dong-hai, LI Ke-fu, ZHAO Si-feng. (2015). Study on law and its influence factors of three main jujube disease in south xinjiang under dwarf and close-planting managements [J].NorthernHorticulture, (3):105-108. (in Chinese)

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      [38]蔣黎艷,趙其陽,龔蕾,等.超高效液相色譜串聯(lián)質譜法快速檢測柑橘中的5種鏈格孢霉毒素[J].分析化學,2015,(12): 1 851-1 858.

      JIANG Li-yan, ZHAO Qi-yang, GONG Lei, et al. (2015). Rapid Determination of fiveAlternariaMycotoxins in citrus by ultra-high performance liquid chromatography-tandem mass spectrometry [J].AnalyticalChemistry, (12):1,851-1,858. (in Chinese)

      Mycotoxin Analysis of Main PathogenAlternariaalternateof Jujube Fruit Shrink and Determination of Black Spot Disease Toxin Content in Xinjiang

      HE Li,GUO Kai-fa,Ainiguli Yiming,ZHAO Si-feng

      (KeyLaboratoryforOasisAgriculturalPestManagementandPlantResourceUtilizationatUniversitiesofXinjiangUygurAutonomousRegion/CollegeofAgronomy,ShiheziUniversity,ShiheziXinjiang832003,China)

      【Objective】 In order to make clear mycotoxins species and content of mainly pathogenAlternariaalternateof jujube fruit shrink, jujube black spot, and mycotoxins species and content of jujube black spot diseased fruits, this study aims to provide the basis for the safety evaluation of jujube processing and consumption in Xinjiang.【Method】Utra-performance liquid chromatography-tandem mass spectrometry was established as the method to determine the variation and content of mycotoxins from 10Alternariastrains and the onset fruit of jujube black spot in different severities and the species and content of toxin in artificially inoculated fruits.【Result】The results showed that 10Alternariastrains could produce 4 kinds ofAlternariamycotoxins: alternariol (AOH), alternariol monomethylether (AME), altenuene(ALT) and tenuazonic acid (TeA).TeA, ALT, AME and AOH were detected in different disease onset of jujube fruit which were affected byA.alternateand artificial cultivation conditions, the highest content of TeA, range of 3.1×103-5.5×103mg/kg; AME, AOH and ALT were 7.2×102-6.4×102mg/kg, 1.2-3.8×102mg/kg and 0.09-5.08 mg/kg. After inoculation withA.alternaria, alarge number of TeA were not only detected injuring jujube fruit, but also detected in the uninjured jujube fruit. Not only the toxin was detected in the onset jujube fruit, but also a large amount of toxin was found in the jujube without injury, although there was a small amount ofAlternariatoxin in healthy jujube fruit.【Conclusion】The main pathogens (Alternariaalternate) leading to jujube fruit shrink and jujube black spot in Xinjiangcan produce a lot ofAlternariamycotoxins, which have such great influence on the production and value of Xinjiang red jujube that it was inedible. Therefore, it should be paid more attention to in fresh jujube processing and risk assessment, and to reduce the infection of pathogens in jujube planting and storage, so as to avoid more pollution. So in jujube planting and storage period, the pathogen infection must be reduced in order to avoid more contamination.

      jujube; short ultivars; growth stages; major gene plus polygene inheritance; genetic analysis

      ZHAO Si-feng (1975- ), male, native place: Bazhong Sichuan. Professor, PhD. research field: Biological control of plant pest. (E-mail)Zhsf_agr@shzu.edu.cn

      10.6048/j.issn.1001-4330.2017.06.014

      2017-03-09

      國家星火計劃“南疆紅棗主要病蟲害綠色防控技術示范與推廣”(2015GA891007)

      何麗(1991-),女,四川南充人,碩士研究生,研究方向為植物病理學,(E-mail)602157044@qq.com

      趙思峰(1975-),男,教授,博士,博士生導師,研究方向為植物病蟲害生物防治,(E-mail)zhsf_agr@shzu.edu.cn

      S432.4+2

      A

      1001-4330(2017)06-1076-09

      Supported by: The National Spark Program of China "Demonstration and Popularization of Green Prevention and Control Technology for Main Diseases and Pests of Jujube in Southern Xinjiang"(2015GA891007)

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