葡萄连作障碍机理及调控途径的研究
摘要
葡萄连作后植株长势衰弱、病害蔓延,严重制约着葡萄产业的发展,为了探讨葡萄连作障碍产生的原因,本试验以连作葡萄30年和种植葡萄3年的葡萄园土壤为研究对象,以晚红葡萄(Vitis vinifera L. Red Globe)组培苗、山河二号葡萄(V. amurensis×V. riparia No.2)组培苗及贝达(V. riparia×V labrusca cv. Beta)葡萄盆栽苗为试材,分析了葡萄连作后土壤微生物、理化性质的变化；研究了根系分泌物、浸提液及腐解物的化感作用,确定了主要自毒物质,并深入分析了自毒物质对葡萄植株生长及根际微环境的影响,同时探讨了不同有机物料及菌剂对葡萄连作障碍的缓解作用,为连作障碍的综合防治提供了理论依据。主要结果如下：
1.生物因素是导致葡萄连作障碍的重要原因之一。连作园根区土盆栽葡萄,植株根系活力下降,叶片的叶绿素含量及净光合速率降低,植株长势衰弱。连作土灭菌后葡萄叶片SOD活性增强,MDA含量降低,根系活力随生长期的延长,表现出先下降再上升的趋势,植株长势明显增强。并且连作土壤灭菌后葡萄根系分泌的还原糖及氨基酸总量显著降低,分别比连作土壤降低了72.60%和91.10%；氨基酸的种类及其含量也发生改变。
2.葡萄连作后根际土壤微生物数量及种群结构发生改变。葡萄连作后根际土壤细菌、放线菌的数量及比例降低,而真菌的数量及比例增加。连作使土壤细菌和真菌的多样性增加,根际土壤的微生物多样性高于非根际土壤。连作后细菌Flavobacterium sp.(DQ339585)(?)Bacillus sp. (AY039821)种群数量减少,Pedobacter sp. (AJ871084)种群数量增多；真菌中Omphalina farinolens(EF413029)出现, Pestalotiopsis sp.(DQ657877, DQ657875, DQ657871)、Phacidium lacerum (DQ470976)和Lecythophora decumbens(AF353597)种群数量减少,Pilidium acerinum voucher (AY48709)种群数量增多。其中Bacillus sp.、Flavobacterium sp.和Pestalotiopsis sp.对病原菌具有颉颃作用,连作园根际土中上述微生物种群数量的减少,不利于葡萄对病原菌的颉颃;Pilidium acerinum voucher数量的增加可能与葡萄连作后病害加重有关。
3.葡萄连作后土壤物理性质未出现恶化,但土壤养分比例发生失调。葡萄连作后并没有增加根区土壤的容重、酸度及盐分含量水平。根区土壤有机质及大量元素随着葡萄种植时间延长而增加,微量元素Fe和Mn含量减少,Zn和Cu含量增加,Zn/Mn、Zn/Fe、N/Fe、N/Mn、P/Fe、P/Mn、Ca/Fe、和Ca/Mn等失调比例较高,其中Zn/Mn、Zn/Fe比例失调最为严重。
4.根系分泌物、浸提液及腐解物具有显著的自毒作用,在它们中确定了5种主要自毒物质。室内组培苗试验结果表明,葡萄根系分泌物、浸提液及根系腐解物具有显著的自毒作用,在它们中分离鉴定出对羟基苯甲酸、水杨酸、香豆酸、苯丙酸和苯甲酸5种酚酸类物质,它们均是影响葡萄生长的重要自毒物质。在根系分泌物和腐解物中推断出5种酸类(邻苯二甲酸、软脂酸、柠檬酸、没食子酸和乌头酸)、2种醇类(1-二十一醇和β-谷甾醇)物质,其中邻苯二甲酸、软脂酸、柠檬酸和没食子酸对葡萄生长表现出“低促高抑”的化感效应,1-二十一醇对葡萄生长的影响无规律性变化,而β-谷甾醇在低浓度时抑制葡萄的生长,高浓度时对葡萄生长具有促进作用。
5.根系分泌物和腐解物能够改变土壤微生物种群结构,影响根际土壤中速效N、P、K水平。根系分泌物处理后,根际土壤碱解N的含量与对照差异不显著,低浓度的根系分泌物处理(E100和E200)增加了根际土壤速效P和速效K含量,高浓度的根系分泌物(E300)处理表现相反的趋势,根系分泌物处理使葡萄根际土壤由“细菌型”向“真菌型”方向转化。根系腐解物处理提高了根际土壤速效N、P、K的含量,降低了根际土壤的真菌种群多样性,使有益的木霉菌消失,而出现了与病害有关的腐霉属和根串珠霉属。
6.化感物质苯丙酸和水杨酸主要是通过干扰植物细胞膜及相关生理过程影响葡萄的生长。盆栽试验表明不同浓度的(0.1 mmo1·L-1,1 mmo1·L-1,10 mmo1·L-1)苯丙酸和水杨酸增加了葡萄植株细胞膜透性、降低了SOD活性及植株根系活力,影响了光合作用和叶片物质积累等生理生化过程,进而使葡萄株高、茎粗、地上鲜重和地下鲜重发生变化,导致植株长势衰弱。
7.稻草和玉米秸秆能够改善连作土壤物理性质,活化土壤营养成分,改善根际微域环境而促进葡萄植株的生长。连作土壤施入稻草和玉米秸秆后,土壤容重降低,孔隙度增加,有机质、速效P和速效K含量均有不同程度的提高,植株长势增强(除连作土中施入0.5%的稻草处理),并且土壤微生物区系的比例发生改变,细菌比值增加。土壤磷酸酶活性随着有机物料腐解时间延长而增强,但土壤过氧化氢酶活性并没有因稻草的施入而提高,玉米秸秆处理显著增加了土壤过氧化氢酶活性。
8.丛枝菌根真菌可以促进连作葡萄生长,改变植株根系分泌特性。连作土灭菌后接种丛枝菌根真菌(Arbuscular Mycorrhizal Fungi, AMF)[地表球囊霉Glomus versiforme(简称G.V)、摩西球囊霉Glomus mosseae (简称GM)和幼套球囊霉Glomus etunicatum(简称G.E)]促进了葡萄植株的生长,叶片SOD活性增强,MDA含量降低,根系活力增强,其中G.V和G.M处理效果最显著。接种菌剂后,葡萄根系分泌的物质种类及其含量也发生变化,其中G.V处理葡萄根系分泌的物质种类较多,并且含有特异性成分橙花叔醇。
9.淡紫拟青霉、木霉菌剂及重茬EB是缓解葡萄连作障碍的重要途径。葡萄连作土壤接种淡紫拟青霉、木霉菌剂及EB后,增强了叶片及根系的SOD活性,降低了MDA含量,叶片的可溶性糖、淀粉及蛋白质含量增加,根系呼吸速率提高,植株长势明显增强。其中淡紫拟青霉中以DZ2(30 g淡紫拟青霉/槽连作土)处理效果最好,木霉菌剂中以MM2(500 g健根宝/槽连作土)处理较好,而EB中以EB1(20 g/槽连作土)处理植株长势最强。综合来看,连作土壤中施入木霉菌剂及EB对葡萄生长的促进效果优于淡紫拟青霉。
The grape replant problem is expressed as stunted growth, low productivity and a decline in tree vigor leading to shortened economic life. At present, the problem is common in all grape growing regions and has seriously restricted the development of grape industry. In order to study the reason of grape replanting, using rhizosphere soil, non-rhizosphere soil and fallow soil of replanted vineyard and new-planted vineyard which had been planted grape for 30 years and 3 years respectively as research object, and tissue culture seedling of Red Globe (V. vinifera L. cv), tissue culture seedling of'Shanhe No.2'grape and pot seedling of Beta (V riparia×V. labrusca cv.) as experimental material, changes of soil microbe and physic-chemical properties were analyzed, and allelopathy of root exudates, root aqueous extracts and root decomposing products were studied, and major allelochemicals were determined, and its effect on grape growth and rhizosphere microenvironment were further analyzed, at the same time, the effect of adding organic materials and inocula into replanting soil on alleviating grape replant obstacle were studied. The major results were as follow:
1. Biological factor played a causal role in the development of grape replant obstacle. Based on pot experiment, the results indicated that root activity, leaves chlorophyll content and net photosynthetic rate which plant grew in root zone soils of replanted vineyard decreased compared to plants which grew in new-planted vineyard, and plants growth was weak. After sterilization of replanting soil, SOD activity of grape leaves increased, while MDA content decresed, and root activity enhanced first and then declined, and plant height, stem diameter, fresh weight of shoot, fresh weight of root increased. Comparing with grape growing on replanting soil, the content of soluble sugar and amino acid of root exudate for grape growing on sterilized replanting soil deceased by 72.60% and 91.10%, and amino acid composition of root exudation also changed.
2. Soil microbial number and population structure changed after grape replanting. The numbers and proportions of bacteria and actinomycetes in rhizosphere soil of replanted vineyard decreased, and the number and proportion of fungi increased. Bacterial and fungal diversity in replanted vineyard was higher than that in new-planted vineyard, and the diversity in rhizosphere soil was greater than that in non-rhizosphere soil. Compared with the new-planted vineyard, grape replanting caused a great change in microbial population of rhizosphere soil. The content of Flavobacterium sp. (DQ339585) and Bacillus sp. (AY039821) decreased, while Pedobacter sp. (AJ871084) increased in bacteria. Omphalina farinolens (EF413029) appeared, and the content of Pestalotiopsis sp. (DQ657877, DQ657875, DQ657871), Phacidium lacerum (DQ470976) and Lecythophora decumbens (AF353597) decreased, while Pilidium acerinum voucher (AY48709) increased in fungi. Among them, Bacillus sp., Flavobacterium sp. and Pestalotiopsis sp. in rhizosphere soil of replanted vineyard had antagonism to pathogen, and the decrease of relative amount reduced resistance against pathogen. Increasing of Pilidium acerinum voucher might relate to severe disease after grape replanting.
3. Soil physical properties were not deteriorated after grape replanting, while nutrient ratio was inbalance. Soil bulk density, acidity and conductivity decreased after grape replant. Soil organic matter and macroelements (N, P, K and available Ca, Mg) increased along with the prolonging of grape planting peroid, available Fe, Mn were deficit in replanted vineyard soils, while available Cu, Zn increased. Zn/Mn, Zn/Fe, N/Fe, N/Mn, P/Fe, P/Mn, Ca/Fe and Ca/Mn were imbalance in the root zone, and among them, Zn/Mn, Zn/Fe were most serious imbalance.
4. Grape root exudates, aqueous extracts, root decomposing products had significant autotoxicity, and in which five allelochemicals were identified. Using the testing methods of tissue culture seedling, the results showed there were significant autotoxicity in grape root exudates, aqueous extracts of grape root, root decomposing products, and in which 5 phenolic acids (p-hydroxybenzoic acid, salicylic acid, coumaric acid, phenylpropionic acid and benzoic acid) were isolation and identification, and they were important allelochemicals. 5 acids (phthalic acid, palmitic acid, citric acid, gallic acid and aconitic acid) and 2 alcohols (1-heneicosanol andβ- sitosterol) was infered in root exudates and root decomposing products. Among them, low-concentration phthalic acid, palmitic acid, citric acid, and gallic acid promoted plant growth, and high concentration of which inhibited plant growth significantly. There were no regular change on grape growth under 1-heneicosanol treatment, and low-concentrationβ- sitosterol inhibited plant growth, while high concentration of which promoted plant growth significantly.
5. Root exudates and root decomposing products changed microbial population structure and available N, P, K content of rhizosphere soil. There were no significant difference in available N content of rhizosphere soil between root exduate treatments and control. Low concentration (E100 and E200) root exudates made available P, available K contents of rhizosphere soil increased. Available P, K content of rhizosphere soil decreased under the treatment of high-concentration (E300) root exudates, which made rhizosphere soil transform from bacterial forms to fungial forms. The treatments of root decomposing products increased available N, available P and available K content of rhizosphere soil, decreased fungal diversity, made Trichoderma disappeared, Pythium, Thielaviopsis, Stilbellales appeared.
6. Phenylpropionic acid and salicylic acid inhibited plant growth through disturbing membrane and related physiological process. Based on pot experiment, the results indicated that phenylpropionic acid and salicylic acid of different concentration (0.1mmol·L-1, 1mmol·L-1, 10mmol·L-1) enhanced cell membrane permeability, decreased SOD activity and root activity, affected photosynthesis and dry matter accumulation of grape leaves. So, they decreased significantly plant height, stem diameter, fresh weight of shoot and root with the rise of concentration, and resulted in growth vigor weak.
7. Rice straw and corn straw improved soil physical properties, activated soil nutrients and ameliorate microenviroment of rhizosphere soil, which promoted grape growth. The measures of adding different proportion rice straw and corn straw into replanted soil decreased soil bulk density, increased soil porosity, organic matter content, available P, available K, and seedlings had strong growth vigor (except for the treatment of 0.5% rice straw adding into replanting soil). The proportion of microflora changed after treatments, and the ratio of bacteria increased. Moreover, soil phosphatase activity increased with decomposition time prolonging of organic materials, and soil catalase activity increased after adding corn straw into replanting soil, while rice straw treatment did not increased soil catalase activity.
8. AMF (arbuscular mycorrhizal fungi) promoted plant growth, and changed root exudation characteristics. AMF (Glomus versiforme G.V, Glomus mosseae G.M and Glomus etunicatum G.E) promoted significantly plant growth, and SOD activity increased, MDA content decresed, root activity increased, among them, GV and G.M treatments were better. AMF (Arbuscular Mycorrhizal Fungi) also affected grape root exudation characteristics, and substance species and relative contents of grape root exudates were changed greatly in different treatment. G.V treatment had more substances and specific component nerolidol.
9. Lilac paecilomyces, biocide and EB were important control approaches of grape replanting. After inoculating lilac paecilomyces, biocide and EB into replanting soil, SOD activity of grape leaves and root activity increased, and MDA content of leaves reduced. Soluble sugar content, starch content and protein content of grape leaves increased after inocula treatment, and root activity raised. Lilac paecilomyces treatment in which DZ2 (30 g lilac paecilomyces of per groove) was best, and biocide treatment in which MM2 (500 g biocide per groove) was best, while EB treatment in which EB1(20 g EB per groove) was best. Comprehensive analysis result showed that the effect of biocide and EB on grape growth better than lilac paecilomyces.
1.生物因素是导致葡萄连作障碍的重要原因之一。连作园根区土盆栽葡萄,植株根系活力下降,叶片的叶绿素含量及净光合速率降低,植株长势衰弱。连作土灭菌后葡萄叶片SOD活性增强,MDA含量降低,根系活力随生长期的延长,表现出先下降再上升的趋势,植株长势明显增强。并且连作土壤灭菌后葡萄根系分泌的还原糖及氨基酸总量显著降低,分别比连作土壤降低了72.60%和91.10%；氨基酸的种类及其含量也发生改变。
2.葡萄连作后根际土壤微生物数量及种群结构发生改变。葡萄连作后根际土壤细菌、放线菌的数量及比例降低,而真菌的数量及比例增加。连作使土壤细菌和真菌的多样性增加,根际土壤的微生物多样性高于非根际土壤。连作后细菌Flavobacterium sp.(DQ339585)(?)Bacillus sp. (AY039821)种群数量减少,Pedobacter sp. (AJ871084)种群数量增多；真菌中Omphalina farinolens(EF413029)出现, Pestalotiopsis sp.(DQ657877, DQ657875, DQ657871)、Phacidium lacerum (DQ470976)和Lecythophora decumbens(AF353597)种群数量减少,Pilidium acerinum voucher (AY48709)种群数量增多。其中Bacillus sp.、Flavobacterium sp.和Pestalotiopsis sp.对病原菌具有颉颃作用,连作园根际土中上述微生物种群数量的减少,不利于葡萄对病原菌的颉颃;Pilidium acerinum voucher数量的增加可能与葡萄连作后病害加重有关。
3.葡萄连作后土壤物理性质未出现恶化,但土壤养分比例发生失调。葡萄连作后并没有增加根区土壤的容重、酸度及盐分含量水平。根区土壤有机质及大量元素随着葡萄种植时间延长而增加,微量元素Fe和Mn含量减少,Zn和Cu含量增加,Zn/Mn、Zn/Fe、N/Fe、N/Mn、P/Fe、P/Mn、Ca/Fe、和Ca/Mn等失调比例较高,其中Zn/Mn、Zn/Fe比例失调最为严重。
4.根系分泌物、浸提液及腐解物具有显著的自毒作用,在它们中确定了5种主要自毒物质。室内组培苗试验结果表明,葡萄根系分泌物、浸提液及根系腐解物具有显著的自毒作用,在它们中分离鉴定出对羟基苯甲酸、水杨酸、香豆酸、苯丙酸和苯甲酸5种酚酸类物质,它们均是影响葡萄生长的重要自毒物质。在根系分泌物和腐解物中推断出5种酸类(邻苯二甲酸、软脂酸、柠檬酸、没食子酸和乌头酸)、2种醇类(1-二十一醇和β-谷甾醇)物质,其中邻苯二甲酸、软脂酸、柠檬酸和没食子酸对葡萄生长表现出“低促高抑”的化感效应,1-二十一醇对葡萄生长的影响无规律性变化,而β-谷甾醇在低浓度时抑制葡萄的生长,高浓度时对葡萄生长具有促进作用。
5.根系分泌物和腐解物能够改变土壤微生物种群结构,影响根际土壤中速效N、P、K水平。根系分泌物处理后,根际土壤碱解N的含量与对照差异不显著,低浓度的根系分泌物处理(E100和E200)增加了根际土壤速效P和速效K含量,高浓度的根系分泌物(E300)处理表现相反的趋势,根系分泌物处理使葡萄根际土壤由“细菌型”向“真菌型”方向转化。根系腐解物处理提高了根际土壤速效N、P、K的含量,降低了根际土壤的真菌种群多样性,使有益的木霉菌消失,而出现了与病害有关的腐霉属和根串珠霉属。
6.化感物质苯丙酸和水杨酸主要是通过干扰植物细胞膜及相关生理过程影响葡萄的生长。盆栽试验表明不同浓度的(0.1 mmo1·L-1,1 mmo1·L-1,10 mmo1·L-1)苯丙酸和水杨酸增加了葡萄植株细胞膜透性、降低了SOD活性及植株根系活力,影响了光合作用和叶片物质积累等生理生化过程,进而使葡萄株高、茎粗、地上鲜重和地下鲜重发生变化,导致植株长势衰弱。
7.稻草和玉米秸秆能够改善连作土壤物理性质,活化土壤营养成分,改善根际微域环境而促进葡萄植株的生长。连作土壤施入稻草和玉米秸秆后,土壤容重降低,孔隙度增加,有机质、速效P和速效K含量均有不同程度的提高,植株长势增强(除连作土中施入0.5%的稻草处理),并且土壤微生物区系的比例发生改变,细菌比值增加。土壤磷酸酶活性随着有机物料腐解时间延长而增强,但土壤过氧化氢酶活性并没有因稻草的施入而提高,玉米秸秆处理显著增加了土壤过氧化氢酶活性。
8.丛枝菌根真菌可以促进连作葡萄生长,改变植株根系分泌特性。连作土灭菌后接种丛枝菌根真菌(Arbuscular Mycorrhizal Fungi, AMF)[地表球囊霉Glomus versiforme(简称G.V)、摩西球囊霉Glomus mosseae (简称GM)和幼套球囊霉Glomus etunicatum(简称G.E)]促进了葡萄植株的生长,叶片SOD活性增强,MDA含量降低,根系活力增强,其中G.V和G.M处理效果最显著。接种菌剂后,葡萄根系分泌的物质种类及其含量也发生变化,其中G.V处理葡萄根系分泌的物质种类较多,并且含有特异性成分橙花叔醇。
9.淡紫拟青霉、木霉菌剂及重茬EB是缓解葡萄连作障碍的重要途径。葡萄连作土壤接种淡紫拟青霉、木霉菌剂及EB后,增强了叶片及根系的SOD活性,降低了MDA含量,叶片的可溶性糖、淀粉及蛋白质含量增加,根系呼吸速率提高,植株长势明显增强。其中淡紫拟青霉中以DZ2(30 g淡紫拟青霉/槽连作土)处理效果最好,木霉菌剂中以MM2(500 g健根宝/槽连作土)处理较好,而EB中以EB1(20 g/槽连作土)处理植株长势最强。综合来看,连作土壤中施入木霉菌剂及EB对葡萄生长的促进效果优于淡紫拟青霉。
The grape replant problem is expressed as stunted growth, low productivity and a decline in tree vigor leading to shortened economic life. At present, the problem is common in all grape growing regions and has seriously restricted the development of grape industry. In order to study the reason of grape replanting, using rhizosphere soil, non-rhizosphere soil and fallow soil of replanted vineyard and new-planted vineyard which had been planted grape for 30 years and 3 years respectively as research object, and tissue culture seedling of Red Globe (V. vinifera L. cv), tissue culture seedling of'Shanhe No.2'grape and pot seedling of Beta (V riparia×V. labrusca cv.) as experimental material, changes of soil microbe and physic-chemical properties were analyzed, and allelopathy of root exudates, root aqueous extracts and root decomposing products were studied, and major allelochemicals were determined, and its effect on grape growth and rhizosphere microenvironment were further analyzed, at the same time, the effect of adding organic materials and inocula into replanting soil on alleviating grape replant obstacle were studied. The major results were as follow:
1. Biological factor played a causal role in the development of grape replant obstacle. Based on pot experiment, the results indicated that root activity, leaves chlorophyll content and net photosynthetic rate which plant grew in root zone soils of replanted vineyard decreased compared to plants which grew in new-planted vineyard, and plants growth was weak. After sterilization of replanting soil, SOD activity of grape leaves increased, while MDA content decresed, and root activity enhanced first and then declined, and plant height, stem diameter, fresh weight of shoot, fresh weight of root increased. Comparing with grape growing on replanting soil, the content of soluble sugar and amino acid of root exudate for grape growing on sterilized replanting soil deceased by 72.60% and 91.10%, and amino acid composition of root exudation also changed.
2. Soil microbial number and population structure changed after grape replanting. The numbers and proportions of bacteria and actinomycetes in rhizosphere soil of replanted vineyard decreased, and the number and proportion of fungi increased. Bacterial and fungal diversity in replanted vineyard was higher than that in new-planted vineyard, and the diversity in rhizosphere soil was greater than that in non-rhizosphere soil. Compared with the new-planted vineyard, grape replanting caused a great change in microbial population of rhizosphere soil. The content of Flavobacterium sp. (DQ339585) and Bacillus sp. (AY039821) decreased, while Pedobacter sp. (AJ871084) increased in bacteria. Omphalina farinolens (EF413029) appeared, and the content of Pestalotiopsis sp. (DQ657877, DQ657875, DQ657871), Phacidium lacerum (DQ470976) and Lecythophora decumbens (AF353597) decreased, while Pilidium acerinum voucher (AY48709) increased in fungi. Among them, Bacillus sp., Flavobacterium sp. and Pestalotiopsis sp. in rhizosphere soil of replanted vineyard had antagonism to pathogen, and the decrease of relative amount reduced resistance against pathogen. Increasing of Pilidium acerinum voucher might relate to severe disease after grape replanting.
3. Soil physical properties were not deteriorated after grape replanting, while nutrient ratio was inbalance. Soil bulk density, acidity and conductivity decreased after grape replant. Soil organic matter and macroelements (N, P, K and available Ca, Mg) increased along with the prolonging of grape planting peroid, available Fe, Mn were deficit in replanted vineyard soils, while available Cu, Zn increased. Zn/Mn, Zn/Fe, N/Fe, N/Mn, P/Fe, P/Mn, Ca/Fe and Ca/Mn were imbalance in the root zone, and among them, Zn/Mn, Zn/Fe were most serious imbalance.
4. Grape root exudates, aqueous extracts, root decomposing products had significant autotoxicity, and in which five allelochemicals were identified. Using the testing methods of tissue culture seedling, the results showed there were significant autotoxicity in grape root exudates, aqueous extracts of grape root, root decomposing products, and in which 5 phenolic acids (p-hydroxybenzoic acid, salicylic acid, coumaric acid, phenylpropionic acid and benzoic acid) were isolation and identification, and they were important allelochemicals. 5 acids (phthalic acid, palmitic acid, citric acid, gallic acid and aconitic acid) and 2 alcohols (1-heneicosanol andβ- sitosterol) was infered in root exudates and root decomposing products. Among them, low-concentration phthalic acid, palmitic acid, citric acid, and gallic acid promoted plant growth, and high concentration of which inhibited plant growth significantly. There were no regular change on grape growth under 1-heneicosanol treatment, and low-concentrationβ- sitosterol inhibited plant growth, while high concentration of which promoted plant growth significantly.
5. Root exudates and root decomposing products changed microbial population structure and available N, P, K content of rhizosphere soil. There were no significant difference in available N content of rhizosphere soil between root exduate treatments and control. Low concentration (E100 and E200) root exudates made available P, available K contents of rhizosphere soil increased. Available P, K content of rhizosphere soil decreased under the treatment of high-concentration (E300) root exudates, which made rhizosphere soil transform from bacterial forms to fungial forms. The treatments of root decomposing products increased available N, available P and available K content of rhizosphere soil, decreased fungal diversity, made Trichoderma disappeared, Pythium, Thielaviopsis, Stilbellales appeared.
6. Phenylpropionic acid and salicylic acid inhibited plant growth through disturbing membrane and related physiological process. Based on pot experiment, the results indicated that phenylpropionic acid and salicylic acid of different concentration (0.1mmol·L-1, 1mmol·L-1, 10mmol·L-1) enhanced cell membrane permeability, decreased SOD activity and root activity, affected photosynthesis and dry matter accumulation of grape leaves. So, they decreased significantly plant height, stem diameter, fresh weight of shoot and root with the rise of concentration, and resulted in growth vigor weak.
7. Rice straw and corn straw improved soil physical properties, activated soil nutrients and ameliorate microenviroment of rhizosphere soil, which promoted grape growth. The measures of adding different proportion rice straw and corn straw into replanted soil decreased soil bulk density, increased soil porosity, organic matter content, available P, available K, and seedlings had strong growth vigor (except for the treatment of 0.5% rice straw adding into replanting soil). The proportion of microflora changed after treatments, and the ratio of bacteria increased. Moreover, soil phosphatase activity increased with decomposition time prolonging of organic materials, and soil catalase activity increased after adding corn straw into replanting soil, while rice straw treatment did not increased soil catalase activity.
8. AMF (arbuscular mycorrhizal fungi) promoted plant growth, and changed root exudation characteristics. AMF (Glomus versiforme G.V, Glomus mosseae G.M and Glomus etunicatum G.E) promoted significantly plant growth, and SOD activity increased, MDA content decresed, root activity increased, among them, GV and G.M treatments were better. AMF (Arbuscular Mycorrhizal Fungi) also affected grape root exudation characteristics, and substance species and relative contents of grape root exudates were changed greatly in different treatment. G.V treatment had more substances and specific component nerolidol.
9. Lilac paecilomyces, biocide and EB were important control approaches of grape replanting. After inoculating lilac paecilomyces, biocide and EB into replanting soil, SOD activity of grape leaves and root activity increased, and MDA content of leaves reduced. Soluble sugar content, starch content and protein content of grape leaves increased after inocula treatment, and root activity raised. Lilac paecilomyces treatment in which DZ2 (30 g lilac paecilomyces of per groove) was best, and biocide treatment in which MM2 (500 g biocide per groove) was best, while EB treatment in which EB1(20 g EB per groove) was best. Comprehensive analysis result showed that the effect of biocide and EB on grape growth better than lilac paecilomyces.
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