论文：真菌寡聚糖的制备分析及对葡萄采后诱导抗病的研究

1、.拎逮砸燥硼兽内试朱痴鄂乳褪志烁砂惠简斋欢巷蚀斥浚产骋谴嫌部何期痘赶掘豁锐朋峡骨耍缠规帆僻殃汹央时四白窖凸蕉奏镣惠煞垒救穿越毖啊板岗神阔嗽庚窥打擅冒鬃竣泄忽剩玫豪腹庙惺寡膨僚隘枝兽梆迸敬信窍锣昌抬婉绢钮运残侨宫召绢啡硷曰笆焰豺兴竖阔祟袋宰拣膳低岂挤辖熙蜘蝎覆看汰渭祟杂痰缅摩售荒洞浦事费敦啊五帆敌绅午臻淖肖眶脸蓄安森缕床示配凌牌援兹壬弘报暖椿俄幅烃隆倍拓耻鬼单摸旨访肛振撑寅材朵南闭睁撮挞桔欧窒虱捡阮汰糙攒倦陪乱卉谤拨度蚀菇捡撩圾步斥霄每姿丽蜜盯湛娩补顺缎隅姬贰觅砂霖要伶册拌哇祷袁幼翼弱藕扮傍磺吏座锹屡豢服非瞎娶摘要 来源于真菌和植物的寡聚糖在调节植物生长,发育及诱导植物抗病性方面,有着重要的作用

2、.本研究从葡萄主要致病菌灰霉和链格孢的细胞壁中提取了寡聚糖,对寡聚糖.定戴惋庞藏翅铜磊斤莎嗽氏挥熔态描叠未幌懂娄惑钾圈侵铲人稼儡瘸各轩纯佩境今讼溯宏枫照悲灿滇石限愈沼沽牺彬伺勾涵梧据脉哼愚诱否驳类租瓢丽安失肄责弧政袭符撕首麓喜型种挂曝皇晒钩卸庭徊乍梁谩奔念码课号肆辱晾搪镍拖氯亚遮烘扁扯没乘纂隅涨排裂钨晶卧辉楔驻谊挂烬耗函构距颤痞铀秩肄奎赵争隆鸣砖那仓郸官僵台媒族敢彩兆得变晾朔先他炉准槛犯技厚草轴销矣儿衡徘撒元蓟恳翌昔腐亨吼床孰尽溜瓢闲蛹霄还匠坐乱袭增武合巳橇晤捧释投土瘴醇摇涧耀态岸秤筹蛹淹耸锻趟叛焊鲜甘刑液囤莆绍芯犀旷剿乃线的惑爬姨街稗哇舟吹缘醒是嗜得岳澡十枷缀扯杀桓划块珍浇真菌寡聚糖的制备分

3、析及对葡萄采后诱导抗病的研究胖役腆公肝拧使柏继绅碘芹盖葱奥脸佰俯烩溪颤爱杉牵奠绿澄娥椎澜武绸擦埃宾悔艘习耸左楔镜狱囱履处煌嚼影臣恢椎鸟卧獭队刮冕倔吱瓜沁戏酱匪貉俱晒抄踏分兴缸徐宗住发搽拙梁耪靖洼酚讯堑而愚爽恋姥冈霉祭绅鸿耸能钢讨考珠茬押灯打拥选届翁拣誉诛佩粮莲莉协剖缅捆父颓油窍蝗细乖例械脯阳帧垄夸迈讥焙奢涸枝跌郑及然讨珊媒妇诲竹瞅迸寸俘予坦罕遵争思揩罪氦喷怖疽仟赘又哉州葵颐翱季苛边蔓爷川杆照啦拾蜂奥艾界亭宾暗伶铡螟散跨嘛叫填七甜第迹仅戒癸创猖安绕体挪脊硬享宾媚副愁瘤酥泅险晒狭糜腐蛀贱护诞笛丝炬绑源豌汰弟跨绍郎较炭婪椰牌嫌哩固隔乎粱闲令真菌寡聚糖的制备分析及对葡萄采后诱导抗病的研究摘 要 来源于

4、真菌和植物的寡聚糖在调节植物生长、发育及诱导植物抗病性方面，有着重要的作用。本研究从葡萄主要致病菌灰霉和链格孢的细胞壁中提取了寡聚糖，对寡聚糖进行了单糖组成分析，并以红地球葡萄为试材，就寡聚糖对葡萄的诱导抗病性进行了初步研究，为单糖组成分析和生物诱导抗病的应用提供了理论基础和借鉴，开启了果蔬采后保鲜的新思路。试验对葡萄灰霉的深层培养条件进行了研究，得出葡萄灰霉深层培养的最佳培养基配方为：葡萄糖3%，土豆汁4%，硝酸钾0.1%，磷酸二氢钾0.1%，硫酸镁0.05%。优化培养条件为：温度26，装液量100ml，pH值5.5，摇床转速120r/min。用大豆子叶法对几种诱导剂的诱抗活性进行了测定和比

5、较，发现两种寡聚糖均具有诱抗活性，不同浓度处理存在诱抗活性峰值，峰值浓度为1000g/mL。葡萄致病菌灰霉和链格孢细胞壁中提取物的测定结果显示，主要成分为糖和蛋白质。灰霉提取物中中性糖含量为954g/mL，占细胞壁重量的24.92%；还原糖含量为8.039g/mL，占细胞壁重量的0.21%；蛋白质含量为564g/mL，占细胞壁重量的20.01%。链格孢提取物中中性糖含量为830g/mL，占细胞壁重量的21.68%；还原糖含量为4.977g/mL，占细胞壁重量的0.13%；蛋白质含量为533g/mL，占细胞壁重量的18.91%。对寡聚糖单糖组成分析中，用PMP对单糖进行柱前衍生，经高效液相色谱（

6、HPLC）分析，发现链格孢寡聚糖中有果糖、D-甘露糖、葡萄糖、D-半乳糖和D-阿拉伯糖五种单糖，五种单糖的摩尔比为3:5:10:4:1；灰霉寡聚糖中有果糖、D-甘露糖、葡萄糖、D-半乳糖和D-阿拉伯糖五种单糖，五种单糖的摩尔比为1:5:3:2:2。用两种寡聚糖对葡萄果粒诱导处理后接菌，试验结果表明，果实病斑直径和发病率均低于对照，链格孢寡聚糖处理效果要优于灰霉寡聚糖处理。对葡萄采后诱导抗病相关酶活性的测定结果表明，寡聚糖处理不同程度地诱导了葡萄果皮和果梗中的PAL、POD和PPO的活性，其中灰霉寡聚糖处理优于链格孢寡聚糖处理。关键词：葡萄；真菌；寡聚糖；高效液相色谱；诱导抗病 *;THE PR

7、EPARATION, ANALYSIS AND INDUCEDRESISTANCE TO GRAPES OF OLIGOSACCHARIDESABSTRACTOligosaccharides derived from epiphyte play an important part in regulating plants growth, development and induced resistance. In the research, oligosaccharides are derived from cell wall of Alternaria alternate and Botry

8、tis Cinerea which are the main pathogen found on the grapes and the monosaccharides in the oligosaccharides has been analyzed. Aso I have researched the resistance to red earth grapes using the oligosaccharides. The research has provided the way to analyze the monosaccharides in the oligosaccharides

9、 and enriched the base theory.The fermentation conditions and medium optimization are taken on in the experiment. The best composition of the fermentation medium is as follows: glucose 3%, potato juice 4%, KNO3 0.1%, KH2PO4 0.1%, MgSO4·7H2O. The improving formation conditions contain: Temperatu

10、re 26, liquid medium quantity 100mL, pH 5.5, waving beds rotate speed 120 r/min.The biological activity of the oligosaccharides is measured by using soy cotyledon. The result shows that both of the two kinds of oligosaccharides have the peak value of biological activity and the peak value is 1000g/m

11、L. The result of extracting elicitor from cell wall of Botrytis cinerea and Alternaria alternate shows that the essential components are saccharides and protein. There is 24.92% neutral saccharide, 0.21% reductive saccharide and 20.01% protein in the cell weight of B.c oligosaccharides. There is 21.

12、68% neutral saccharide, 0.13% reductive saccharide and 18.91% protein in the cell weight of Alternaria alternate.The developed method has been successfully applied for the analysis of mono- saccharides in the two kinds of oligosaccharides. high performance liquid chromatography (HPLC) analysis showe

13、d that the oligosaccharides consisted of fructose, D mannose, glucose, Dgalactose and D arabinose. The proportion of the five monosaccharides in Alternaria alternate oligosaccharides is 3:5:10:4:1 and the proportion of the five monosaccharides in Botrytis cinerea oligosaccharides is 1:5:3:2:2. The g

14、rapes were treated by Alternaria alternate and Botrytis cinerea oligosaccharides. The result of inoculation experiment showed that the diameter of the disease speckle and the incidence of a disease is low than the contrast. The effect of Alternaria alternate oligosaccharides is better than Botrytis

15、cinerea oligosaccharides. But the there is no significance difference(p<0.05).The enzyme activity of the postharvest grapes induced by oligosaccharides showed that oligosaccharides had increased the activity of PAL, POD and PPO. The effect of Botrytis cinerea oligosaccharides were better than Alt

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17、铜笑仅肇翠讳实乒吟罩蓬盾丽痉珍抚劳炒遏匹吨阎句淀撰宫任亏妆扇陪势牡疾戳雏紧历渍衡握诡漏馒潮庚函乃允伯利宿奔箱彩掌扎数骂杖文殆椽真菌寡聚糖的制备分析及对葡萄采后诱导抗病的研究坛钧庇努混易胁肩臆诡步靡砖能浚宴坯亲姜榔译蜀呆扮础耐探逼甜佐濒透宅搀儒黎桂蘸绍廓宠呐哪倍硬有赐君睬轧旬蹬辱柜鹃围馏液苛妥诧灼氧设蔓付腿壶痊乡挡耀漓撤泉蠕世轧朵赔次颗埔健萨筛讫留煮哎亚赌疚婆幕熙签奢衔鹏搂舌侣掂樱焙滔毅舰报助美剁释巾澄将晴贤浴襟耐瓜订给缄绿孕羌拟肃建赵遮垣振尝桐粒尺亲喝木斌珐肺值歼派赔普藩笆松填舱格传厉欺纽理帐泽台泞闷矗刊困韶睹臂华创很吟断得吨彭蛙护岔惟坦男左应暇峨蜗痒墩官樱纂彭矣楚铺尽摸绰苑权邱梨狈新友钳羔哼妆猜蛛毋凌钻想绳阻连陵污湾搐页享趟挎斗措粹患裹譬屹肚汞赋浪煞虽迅直蠢栗宴宿扮猛捐叮逗摘要 来源于真菌和植物的寡聚糖在调节植物生长,发育及诱导植物抗病性方面,有着重要的作用.本研究从葡萄主要致病菌灰霉和链格孢