亚洲免费av电影一区二区三区,日韩爱爱视频,51精品视频一区二区三区,91视频爱爱,日韩欧美在线播放视频,中文字幕少妇AV,亚洲电影中文字幕,久久久久亚洲av成人网址,久久综合视频网站,国产在线不卡免费播放

        ?

        新疆紅棗棗果主要致病菌交鏈格孢菌產(chǎn)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(nóng)業(yè)科學(xué) 2017年6期
        關(guān)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(nóng)業(yè)病蟲害治理與植保資源利用自治區(qū)高校重點(diǎn)實驗室/石河子大學(xué)農(nóng)學(xué)院,新疆石河子 832003)

        新疆紅棗棗果主要致病菌交鏈格孢菌產(chǎn)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(nóng)業(yè)病蟲害治理與植保資源利用自治區(qū)高校重點(diǎn)實驗室/石河子大學(xué)農(nóng)學(xué)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(chǎn)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(jù)?!痉椒ā坎捎贸咝б合嗌V-串聯(lián)質(zhì)譜法,測定10株鏈格孢菌產(chǎn)生的毒素種類及含量,并對棗果黑斑病不同嚴(yán)重度發(fā)病棗果和人工接種果實中毒素種類及含量進(jìn)行測定。【結(jié)果】10株交鏈格孢供試菌株可產(chǎn)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(xì)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(nèi)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(jié)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(chǎn)生鏈格孢霉毒素,且產(chǎn)毒量高,危害嚴(yán)重,影響了新疆紅棗的產(chǎn)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(fēng)險評估中應(yīng)引起關(guān)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(dǎo)致多種農(nóng)作物病害,同時該屬真菌還可以產(chǎn)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(xì)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]?!厩叭搜芯窟M(jìn)展】目前已在蘋果、柑橘、番茄、土豆、小麥、葵花籽等農(nóng)產(chǎn)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(rèn)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(guān)。AOH能引起食道上皮細(xì)胞增殖,并能引起胚胎食道鱗狀細(xì)胞癌變,用AOH處理老鼠皮下,可誘導(dǎo)鱗狀上皮細(xì)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(dǎo)NIH 3T3細(xì)胞轉(zhuǎn)化,轉(zhuǎn)化后的細(xì)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(qiáng),具多種生物特性,如抗病毒[13]、抗腫瘤、抑菌[14]、細(xì)胞毒性、植物毒性[15],抑制蛋白質(zhì)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(guān)[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(jìn)行風(fēng)險發(fā)布,其中TeA已列入美國食品藥物管理局(FDD)有毒化學(xué)物質(zhì)登記冊?!颈狙芯壳腥朦c(diǎn)】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(chǎn)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。【擬解決的關(guān)鍵問題】研究采用超高效液相色譜-串聯(lián)質(zhì)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(jìn)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(fēng)險評估提供依據(jù)。

        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(chǎn)區(qū)的棗縮果病和棗果黑斑病病樣上,經(jīng)形態(tài)學(xué)和分子生物學(xué)方法鑒定為交鏈格孢菌(A.alternate),菌種保藏于石河子大學(xué)新疆綠洲農(nóng)業(yè)病蟲害治理與植保資源利用自治區(qū)高校重點(diǎn)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(jìn)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(nèi)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(biāo)準(zhǔn)品購買于美國Sigma公司;細(xì)交鏈格孢菌酮酸(TeA,純度>98%)和交鏈孢烯(ALT,純度>98%)標(biāo)準(zhǔn)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(zhì)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(zhì)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(zhì)譜掃描方式:多反應(yīng)監(jiān)測(MRM);毛細(xì)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(zhì)譜參數(shù)。表1

        表1 4種鏈格孢霉毒素的串聯(lián)質(zhì)譜測定參數(shù)
        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(xì)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(fù),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(fù)中取1 g菌餅,用液氮研磨充分,于2 mL離心管中。離心管內(nèi)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(zhuǎn)入一個新2 mL離心管內(nèi),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(jī)濾膜,濾液經(jīng)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(shù)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(zhuǎn)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(jī)濾膜,濾液經(jīng)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(jìn)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(nèi)(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(zhǔn)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(biāo),每個水平重復(fù)測定6次,按1.3.2樣品處理方法處理并測定,以進(jìn)樣質(zhì)量濃度X(ng/mL)為橫坐標(biāo)、峰面積Y為縱坐標(biāo)繪制標(biāo)準(zhǔn)曲線。

        2 結(jié)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(zhì)量濃度為100 ng/mL的混標(biāo)溶液(AOH、AME、ALT和TeA混合物)在3 min內(nèi)達(dá)到有效分離且峰型良好。保留時間為0.81和2.3 mim的物質(zhì)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(zhì)譜連用技術(shù),結(jié)合濃度為100 ng/mL標(biāo)準(zhǔn)溶液中4種鏈格孢霉毒素(AOH、AME、ALT和TeA)的MRM定量離子對質(zhì)譜圖,以離子比為參照確定混合物中組分,并不影響試驗結(jié)果,試驗可行。圖1,圖2

        圖1 4種鏈格孢霉毒素的混合標(biāo)準(zhǔn)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(nèi)均有良好的線性關(guān)系,R2>0.99,平均回收率在70%~95%,相對標(biāo)準(zhǔn)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(chǎn)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(dá)5.39×103mg/kg,hm-16產(chǎn)毒最低,含量為35.9 mg/kg;菌株92-3-1和hm-24產(chǎn)生的毒素種類最多,4種鏈格孢霉毒素均可產(chǎn)生;菌株30-HB-2和36-3-3產(chǎn)生毒素種類最少,僅產(chǎn)生TeA;菌株hm-16、hm-31和hm-35可產(chǎn)生AOH、AME、ALT,產(chǎn)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(biāo)準(zhǔn)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(zhì)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(chǎn)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(dá)3.1×103mg/kg,AME、AOH、ALT分別可達(dá)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(yùn)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(nèi)均產(chǎn)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(chǎn)毒量最高,TeA為2 102.90 mg/kg,AME為1 390.86 mg/kg;hm-16產(chǎn)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(chǎn)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(yùn)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(yīng)性強(qiáng),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(nóng)作物和經(jīng)濟(jì)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?shù)葒邪l(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(yán)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(jīng)濟(jì)損失,發(fā)生嚴(yán)重時可使梨采后損失達(dá)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(jié)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(yùn)用超高液相色譜質(zhì)譜串聯(lián)技術(shù)測定了小麥、番茄、核桃、藍(lán)莓中的鏈格孢霉毒素,結(jié)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(jié)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(jīng)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(nóng)產(chǎn)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(chǎn)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(shù)的棗果作為研究對象,10株供試菌株均能產(chǎn)生鏈格孢菌毒素,不同菌株產(chǎn)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(chǎn)生4種毒素,且產(chǎn)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(yùn)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(chǎn)生的毒素。

        4 結(jié) 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(zhì)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(chǎn)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(chǎn)生量最高,最高可達(dá)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(shù)的棗果黑斑病果實進(jìn)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(guān)鍵問題,采收時應(yīng)當(dāng)將健康果實與病果分開,防止健康果實被病果污染。

        References)

        [1] Logrieco, A., Bottalico, A., Mulé, G., Moretti, A., & Perrone, G. (2003). Epidemiology of toxigenic fungi and their associated mycotoxins for some Mediterranean crops.EuropeanJournalofPlantPathology, 109(7): 645-667.

        [2] Thomma, B. P. H. J. (2003). Alternaria, spp.: from general saprophyte to specific parasite.MolecularPlantPathology, 4(4): 225-236.

        [3] Patriarca, A., Azcarate, M. P., Terminiello, L., & Fernández, P. V. (2007). Mycotoxin production by alternaria strains isolated from argentinean wheat.InternationalJournalofFoodMicrobiology, 119(3): 219-222.

        [4] Janardhanan, K. K., & Husain, A. (1983). Studies on isolation, purification and identification of tenuazonic acid, a phytotoxin produced by alternaria alternata, (fr.) keissler causing leaf blight of datura innoxia, mill.Mycopathologia, 83(3): 135-140.

        [5] Logrieco, A., Moretti, A., & Solfrizzo, M. (2009). Alternaria toxins and plant diseases: an overview of origin, occurrence and risks.WorldMycotoxinJournal, 2(2): 129-140.

        [6] O Ostry, V. (2008). Alternaria mycotoxins: an overview of chemical characterization, producers, toxicity, analysis and occurrence in foodstuffs.WorldMycotoxinJournal, 1(2): 175-188.

        [7] Magnani, R. F., Souza, G. D. D., & Rodrigues-Filho, E. (2007). Analysis of alternariol and alternariol monomethyl ether on flavedo and albedo tissues of tangerines ( citrus reticulata ) with symptoms of alternaria brown spot.JournalofAgricultural&FoodChemistry, 55(13): 4,980-4,986.

        [8] Logrieco, A., Visconti, A., & Bottalico, A. (1990). Mandarin fruit rot caused by alternaria alternata and associated mycotoxins.PlantDisease, 74(6): 415-417.

        [9] Dong, Z. G., Liu, G. T., Dong, Z. M., Qian, Y. Z., An, Y. H., & Miao, J. A., et al. (1987). Induction of mutagenesis and transformation by the extract of alternaria alternata isolated from grains in linxian, china.Carcinogenesis, 8(7): 989-991.

        [10] Liu, G. T., Qian, Y. Z., Zhang, P., Dong, W. H., Qi, Y. M., & Guo, H. T. (1992). Etiological role of alternaria alternata in human esophageal cancer.ChineseMedicalJournal, 105(5): 394-400.

        [11] Lehmann, L., Wagner, J., & Metzler, M. (2006). Estrogenic and clastogenic potential of the mycotoxin alternariol in cultured mammalian cells.Food&ChemicalToxicology, 44(3): 398-408.

        [12] An, Y. H., Zhao, T. Z., Miao, J., Liu, G. T., Zheng, Y. Z., & Xu, Y. M., et al. (1989). Isolation, identification, and mutagenicity of alternariol monomethyl ether.JournalofAgricultural&FoodChemistry, 37(5): 1,341-1,343.

        [13]M Miller, F. A., Rightsel, W. A., Sloan, B. J., Ehrlich, J., French, J. C., & Bartz, Q. R., et al. (1964). Antiviral activity of tenuazonic acid.Nature,200(4913): 1,338-1,339.

        [14] Gitterman, C. O. (1965). Antitumor, cytotoxic, and antibacterial activities of tenuazonic acid and congeneric tetramic acids.JournalofMedicinalChemistry, 8(4): 483-486.

        [15] Lebrun, M. H., Nicolas, L., Boutar, M., Gaudemer, F., Ranomenjanahary, S., & Gaudemer, A. (1988). Relationships between the structure and the phytotoxicity of the fungal toxin tenuazonic acid.Phytochemistry, 27(1): 77-84.

        [16] Shigeura, H. T., & Gordon, C. N. (1963). The biological activity of tenuazonic acid.Biochemistry, 2(2): 1,132-1,137.

        [17]吳春生,馬良,江濤,等.鏈格孢霉毒素細(xì)交鏈格孢菌酮酸的研究進(jìn)展[J].食品科學(xué),2014,35(19): 295-301.

        WU Chun-sheng, MA Liang, JIANG Tao, et al. (2014). A review on tenuazonic acid, a toxic produced byAlternaria[J].FoodScience, 35(19):295-301.(in Chinese)

        [18] Steyn, P. S., & Rabie, C. J. (1976). Characterization of magnesium and calcium tenuazonate from phoma sorghina.Phytochemistry, 15(12): 1,977-1,979.

        [19]董寧,馮宏祖,王蘭,等.南疆駿棗黑斑病癥狀表現(xiàn)及病原菌鑒定[J].植物保護(hù)學(xué)報,2016,43(6): 922-927.

        DONG Ning, FENG Hong-zu, WANG Lang, et al. (2016). The disease symptom and the pathogen identification of jujube black spot in southern Xinjiang [J].JournalofPlantProtection, 52(7):53-58. (in Chinese)

        [20]向征,鐘聰慧,胡軍,等.新疆棗果黑斑病病原鑒定[J].新疆農(nóng)業(yè)科學(xué),2013,50(5): 845-850.

        XIANG Zheng, ZHONG Cong-hui, HU Jun, et al. (2013). Identification of Jujube black spot pathogens in Xinjiang, China [J].XinjiangAgriculturalSciences, 50(5):845-850. (in Chinese)

        [21]馬榮,劉曉琳,梁英梅,等. 棗果黑斑病發(fā)生相關(guān)因素分析及田間藥效試驗[J]. 中國農(nóng)學(xué)通報,2015,(16): 182-189.

        MA Rong, LIU Xiao-lin, LIANG Ying-mei, et al. (2015). Analysis of the influencing factors on disease occurrence of Jujube black pot and field efficacy Test [J].ChineseAgriculturalScienceBulletin, (16):182-189. (in Chinese)

        [22] Andersen, B., Hansen, M. E., & Smedsgaard, J. (2005). Automated and unbiased image analyses as tools in phenotypic classification of small-spored alternaria spp.Phytopathology, 95(9): 1,021-1,029.

        [23]方中達(dá).植病研究方法[M].第三版. 北京:中國農(nóng)業(yè)出版社,1998.

        FANG Zhong-da. (1998).Methodofplantpathologyresearch[M]. Third Edition. Beijing: China Agriculture Press, 1998.(in Chinese)

        [24] Zhao, K., Shao, B., Yang, D., Li, F., & Zhu, J. (2015). Natural occurrence of alternaria toxins in wheat-based products and their dietary exposure in China.PLOSOne. 10(6), e0132019.

        [25] Harteveld, D. O. C., Akinsanmi, O. A., & Drenth, A. (2013). Multiple alternaria, species groups are associated with leaf blotch and fruit spot diseases of apple in australia.PlantPathology, 62(2): 289-297.

        [26] Machacek, J. E., & Greaney, F. J. (2011). The "black-point" or "kernel smudge" disease of cereals.CanadianJournalofResearch, (16): 84-113.

        [27]Y B Kang, Y J Zhang , H J Li , et al. (1999). State of the field of wheat on black point disease (in Chinese) [J].TriticalCrops, 19(2): 58- 60. (in Chinese)

        [28]楊曉平,胡紅菊,王友平,等. 梨黑斑病病原菌的生物學(xué)特性及其致病性觀察[J].華中農(nóng)業(yè)大學(xué)學(xué)報,2009,(12):680-684.

        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)

        [29]謝莉,張鐸,張麗萍,等. 拮抗梨黑斑病菌的鏈霉菌的篩選及鑒定[J].河北師范大學(xué)學(xué)報(自然科學(xué)版),2008,32(4):526-529.

        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)

        [30]嚴(yán)進(jìn),施宗偉,宋福,等. 河北和山東鴨梨果實上鏈格孢菌鑒定[J].植物保護(hù)學(xué)報,2009,36(1) :37-43.

        YAN Jin, SHI Zong-wei, SONG Fu, et al. (2009). Identification ofAlternariaisolates from Ya-pear fruits in Hebei and Shandong provinces [J].JournalofPlantProtection, 36( 1) :37-43. (in Chinese)

        [31]史文景,趙齊陽,張耀海,等. 超高效液相色譜結(jié)合分散液液微萃取同時測定果汁中的7種真菌毒素[C]//. 中國儀器儀表學(xué)會分析儀器分會、中國儀器儀表行業(yè)協(xié)會分析儀器分會、中國儀器儀表學(xué)會檢驗檢疫儀器應(yīng)用技術(shù)分會第三屆中國食品與農(nóng)產(chǎn)品質(zhì)量安全檢測技術(shù)國際論壇暨展覽會論文集.北京: 中國北京雄鷹國際展覽有限公司,2014:13-19.

        SHI Wen-jing, ZHAO Qi-yang, ZHANG Yao-hai, et al. (2014).SimultaneousDeterminationofSevenMycotoxinsinFruitJuicebyUltraPerformanceLiquidchromatographyCombinedwithModifiedDispersiveLiquid-LiquidMicroextraction[C]//. China Instrument Society Analysis Instrument Branch, China Instrument Industry Association Analysis Instrument Branch, China Instrument Society inspection and quarantine instrument application technology branch. The third Chinese food and agricultural products quality and safety testing technology international forum and Exhibition proceedings. Beijing, Beijing Lanneret International Exhibition Co, Ltd. : 13-19. (in Chinese)

        [32]陳月萌,李建華,張靜,等. 高效液相色譜-熒光檢測法同時測定水果中的3種鏈格孢霉毒素[J]. 分析試驗室,2012,(6):70-73.

        CHEN Yue-meng, LI Jian-hua, ZHANF Jing, et al. (2016). Simultaneous determination of threeAlternariasin fruits by HPLC with fluorescence detection [J].ChineseJournalofAnalysisLaboratory, (6):70-73. (in Chinese)

        [33] Andersen, B., Nielsen, K. F., Pinto, V. F., & Patriarca, A. (2015). Characterization of alternaria, strains from argentinean blueberry, tomato, walnut and wheat.InternationalJournalofFoodMicrobiology: (196): 1-10.

        [34] Scott, P. M., Zhao, W., Feng, S., & Lau, P. Y. (2012). Alternaria, toxins alternariol and alternariol monomethyl ether in grain foods in canada.MycotoxinResearch, 28(4):261-266.

        [35]張棟海,李克福,趙思峰.新疆南疆矮化密植棗園三種紅棗病害發(fā)生規(guī)律及其影響因素研究[J].北方園藝,2015,(3):105-108.

        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)

        [36]趙燕,郭慶元,王洪凱,等.新疆紅棗病害種類及田間發(fā)生[J].新疆農(nóng)業(yè)科學(xué), 2015,52(3):511-516,589.

        ZHAO Yan, GUO Qin-yuan, WANG Hong-kai, et al. (2015). Investigation of Chinese jujube diseases and their field occurrence in Xinjiang [J],XinjiangAgriculturalSciences, 52(3):511-516,589. (in Chinese)

        [37]王志霞,孫潔,趙思峰,等.新疆矮化密植棗園紅棗葉斑病病原鑒定[J].中國森林病蟲,2013,32(4):1-5.

        WANG Zhi-xia, SUN Jie, ZHAO Si-feng, et al. (2013). Identification of the pathogenic fungus causing leaf spot ofZizyphusjujubein dwarf and close-planting orchards in Xinjiang [J].ForestPestandDisease, 32(4):1-5. (in Chinese)

        [38]蔣黎艷,趙其陽,龔蕾,等.超高效液相色譜串聯(lián)質(zhì)譜法快速檢測柑橘中的5種鏈格孢霉毒素[J].分析化學(xué),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(shù)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(xué),(E-mail)602157044@qq.com

        趙思峰(1975-),男,教授,博士,博士生導(dǎo)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)

        猜你喜歡
        孢霉鏈格棗果
        不同時期施用不同肥料提高山地棗果維生素C 含量
        被孢霉對土壤養(yǎng)分有效性和秸稈降解的影響*
        “二月蘭葉斑病菌甘藍(lán)鏈格孢生物學(xué)特性觀察實驗”的教學(xué)設(shè)計
        甘肅教育(2021年10期)2021-11-02 06:14:26
        16個棗樹品種棗果營養(yǎng)成分分析
        食用菌工廠化生產(chǎn)防治鏈孢霉技術(shù)探究
        食用菌(2019年4期)2019-08-27 06:34:58
        中國月季上兩株鏈格孢屬真菌的分離鑒定及TeA毒素的測定
        歐盟評估鏈格孢霉毒素在不同人群中的膳食暴露風(fēng)險
        桃色頂孢霉代謝產(chǎn)物對綠豆白粉病的防治作用
        西藏砂生槐莖內(nèi)鏈格孢屬真菌生物學(xué)特性研究
        西藏卷葉黃精莖內(nèi)鏈格孢屬真菌生物學(xué)特性研究
        西藏科技(2015年9期)2015-09-26 12:15:34
        在线a免费观看| 日韩亚洲午夜精品一区二区三区| 少妇我被躁爽到高潮在线影片| 婷婷色婷婷开心五月四| 特级a欧美做爰片第一次| 国产一区免费观看| 国产av91在线播放| 青青草手机在线免费观看视频| 亚洲国产精品成人综合色| 亚洲欧美日韩精品高清| 亚洲AV永久无码精品导航| 国产目拍亚洲精品二区| 国产不卡精品一区二区三区| 国产精品你懂的在线播放| 國产AV天堂| 一级a免费高清免在线| 亚洲国产果冻传媒av在线观看| 欧美人与动牲猛交xxxxbbbb| 麻豆密入视频在线观看| 色噜噜亚洲精品中文字幕| 亚洲av熟女天堂系列| 亚洲V在线激情| 久久精品国产亚洲av一| 综合图区亚洲另类偷窥| 一本一道av无码中文字幕| 精品亚洲少妇一区二区三区| 精品精品国产一区二区性色av| 亚洲av永久无码天堂网小说区| 国产av无码专区亚洲av手机麻豆 | 四虎欧美国产精品| 国产丝袜在线福利观看| 极品少妇xxxx精品少妇偷拍| 18禁高潮出水呻吟娇喘蜜芽| 久久久久久AV无码成人| 亚洲最新精品一区二区| 久久综合狠狠色综合伊人| 久久国产品野战| 日本淫片一区二区三区| 久久久久久九九99精品| 91社区视频在线观看| 日本办公室三级在线看|