热胁迫下丹参迷迭香酸代谢途径关键酶基因的表达研究

为探索热胁迫对丹参迷迭香酸途径关键酶基因表达的影响,采用定量RT-PCR法,以Actin与GAPDH作为内参基因,0~48h叶片cDNA作为模板,对迷迭香酸途径7个关键酶基因PAL、C4H、4CL、TAT、HPPD、HPPR和RAS的表达进行分析。通过试验结果构建出这7个关键酶基因0~48h代谢途径表达图谱。其中,PAL、C4H和RAS受热胁迫影响表达量下降;TAT、4CL和HPPD表达量呈先上升后下降趋势;HPPR表达量前期变化不大,后期呈下降趋势。结果表明热胁迫对迷迭香酸途径关键酶基因表达有极显著影响。该表达时序谱的建立为进一步研究热胁迫与酚酸类成分累积之间的关系奠定了基础。     

盐胁迫对番茄幼苗中乙烯信号转导途径一些基因表达的影响

用不同浓度的NaCl溶液分别处理番茄(Lycopersicon esculentum cv．UC-82)幼苗，研究分别处理12、24和48h后乙烯信号转导途径中编码转录因子基因的表达变化。结果表明，不同浓度的NaCl溶液处理番茄幼苗12h能明显促进Le-EIL1基因的表达；0．08mol／L NaCl溶液处理12和24h都能促进Le-E1L2基因的表达；用0．04和0．08mol／L NaCl处理12h和24h后，Le-EIL3基因表达都得到增强。用0．04和0．08mol／L NaCl溶液处理番茄幼苗12h能显著地促进Le-ERF1基因的表达；用3种浓度的NaCl溶液0．04，0．08和0．16mol／L处理番茄幼苗12h后均能明显的促进Le-ERF2基因的表达；用不同浓度的NaCl溶液分别处理番茄幼苗Pti4基因的表达变化不大。这些结果说明Le-EILs和Le-ERFs家族成员基因在不同浓度NaCl溶液和不同处理时间条件下表现不同的表达模式。     

水稻白叶枯病菌蛋白质激发子HarpinXoo诱导植物的防卫反应

摘要：含质粒pHRF4的表达菌株BLHR4在培养基中经IPTG诱导，16 h Harpin表达接近最高量。用不同浓度的HarpinXoo喷雾处理烟草(Nicotiana tabacum L.)、油菜(Brassica campestris L.)和番茄(Lycopersicon esculentum Mill.)，对烟草花叶病毒(TMV)、油菜菌核病(Sclerotinia sclerotiorum (lib.) de Bary)和番茄灰霉病(Botrytis cinerea Pers.)均表现较好的诱导抗病效果。以浓度为100μg/mL的HarpinXoo处理烟草后，分别测定叶片中与防卫反应相关的一些生理指标的变化，结果表明，处理叶片中苯丙氨酸解氨酶（PAL）、过氧化物酶（PO）和多酚氧化酶（PPO）活性都有不同程度的增强。三种酶活性高峰分别在处理后24、12和3 h出现，分别比清水对照增加110.1%、211.2%和273.0%；同时，两个病程相关蛋白（pathogenesis-related protein，PR）基因PR-1b和PR-1a在HarpinXoo处理24 h后被诱导显著表达，36 h时达到最高表达水平。这表明HarpinXoo与HarpinEa一样能够诱导烟草不同抗病信号通路的启动，从而使植株获得抗病性。     

Preparation of Bioactive Recombinant Chicken Leptin Fusion Protein

将鸡Leptin成熟肽的cDNA片段插入表达载体pRSET A的Nhe Ⅰ和Hind Ⅲ两位点之间，构建重组表达质粒pLep -SCAU2并转化大肠杆菌(Escherichia coli )菌株BL21（DE3）, 将重组菌在含氨苄青霉素100 μg/mL 的LB培养基中培养至OD600大于0.6之后，经IPTG 0.05 mol/L诱导表达出分子量约为17.5 kD的含6×His标记的重组鸡Leptin，诱导4 h后表达量达最高，占总菌体蛋白的25％左右,且以包涵体形式存在。该重组鸡Leptin经50％Ni-NTA树脂纯化和透析复性折叠后分离出可溶性部分。对35日龄的矮脚黄鸡腹腔注射该可溶性重组鸡Leptin（2 mg/kg 体重）；注射后60 min内的采食量与对照组差异不显著 （P > 0.05），但在注射后80~120 min都显著低于对照组（P < 0.05）。Leptin处理公鸡的采食量在注射2 h后逐渐上升并在5.5 h时与对照组公鸡相同。但Leptin处理母鸡的采食量持续受到抑制，在注射后5.5 h时仍然显著低于对照组母鸡（P <0.01）。表明所表达的重组鸡Leptin能显著抑制鸡的采食量（母鸡比公鸡效果更为明显），证明所制备的重组鸡Leptin融合蛋白具有生物学活性。     

硅对水稻防御性关键酶活性的影响及其与抗稻瘟病的关系

通过室内溶液培养试验,研究了硅酸盐对接种稻瘟病菌后水稻的病情指数、植株生物量、过氧化物酶（POD）、多酚氧化酶（PPO）和苯丙氨酸解氨酶（PAL）活性的影响。结果表明,施硅能显著降低水稻稻瘟病的发病率和病情指数,防治效果达60.59%;显著减轻稻瘟病对水稻地上部的危害,干物质量显著高于不施硅处理,但对地下部生长影响不明显;接种后水稻叶片POD活性均升高,在第5 d达到最大值,施硅处理显著低于不施硅处理,与稻瘟病抗性成负相关;水稻叶片PPO活性在接种后第5 d达到峰值,从第4 d开始施硅处理显著高于不施硅处理;接种后水稻叶片的PAL活性快速升高,在第24 h达到最大值,施硅处理显著高于不施硅处理,是其1.44倍。说明硅能提高防御性关键酶的活性,参与水稻本身防卫机制,通过一系列的生理生化作用来增强水稻的抗性。     

Pseudomonas putida GM6多聚磷酸盐激酶(ppk)基因的克隆及表达

以一株高效聚磷菌Pseudomonas putida GM6为研究材料.为获得其多聚磷酸盐激酶（polyphosphate kinase,ppk）基因,并验证该基因在磷酸盐转运系统中的作用,根据已报道的ppk基因保守区域设计引物,从其总DNA中成功扩增到ppk基因的部分片段（约528 bp）.随后采用快速染色体步移方法（Self-formed adaptor PCR,SEFA-PCR）技术扩增片段的上下游基因序列,将三个序列拼接,用OMIGA软件分析其ORFs,推测ppk基因全长为2 220 bp（GenBank accession number DQ133537）.构建的多聚磷酸盐激酶表达菌株E. coli BL21（DE3）/ pET29a-ppk经IPTG诱导后3 h时,明显出现分子量约为81 kDa的表达产物.且表达菌株在12 h时的磷去除率高达80%（对照菌株的磷去除率仅为18%）,远高于已报道的40%的去除率.这表明ppk基因在E. coli中的过量表达,导致了E. coli菌体中poly-P的大量聚集,从而大大去除了培养基中的磷酸盐.     

印度梨形孢诱导烟草对黑胫病的抗性及其机理的初步研究

印度梨形孢(Piriformospora indica)是分离自印度塔尔沙漠的植物根系内生真菌,能够诱导多种植物对生物和非生物逆境产生抗性,目前已作为一种模式真菌用于内生真菌与植物互作关系的研究。为了研究印度梨形孢对烟草(Nicotiana tobacum)抗黑胫病的影响,本研究采用烟草疫霉菌(Phytophthora parasitica var.nicotianae)分别接种根部有印度梨形孢定殖与未定殖的烟草,观察接种后根部有印度梨形孢定殖与未定殖的烟草发病情况,测定抗病相关防御酶苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、多酚氧化酶(PPO)的活性变化及抗病相关基因的表达量,并通过平皿对峙试验研究印度梨形孢对烟草疫霉菌生长的影响。结果表明,接种烟草疫霉菌后,根部有印度梨形孢定殖的烟草发病严重度要显著低于对照;接种烟草疫霉菌后第1~10 d,根部有印度梨形孢定殖的烟草叶片中防御酶PAL、POD和PPO活性高于对照,在第1 d,PAL活性是对照烟草的3.99倍,在第2 d,POD活性是对照烟草的1.5倍,在第4 d,PPO活性是对照烟草的2.66倍;实时荧光定量PCR(Real-time quantitative PCR,q RT-PCR)结果显示,接种烟草疫霉菌后第6~96 h,根部有印度梨形孢定殖的烟草叶片中抗病相关基因PR1b、P450-2、hrs203J、Nm IMSP和P450-1的表达量高于对照;在第12 h,PR1b、P450-2、hrs203J的表达量分别是对照烟草的3.8、15.7和3.1倍,在第48 h,Nm IMSP的表达量是对照烟草的2.6倍,在第72 h,P450-1的表达量是对照烟草的1.5倍;平皿对峙试验结果显示印度梨形孢对烟草疫霉菌的生长没有抑制、重寄生作用。研究结果表明,印度梨形孢可显著提高烟草对黑胫病的抗性,对黑胫病的抗性不是通过印度梨形孢分泌的抗菌物质实现的,是与烟草中抗病相关基因表达量的上升、防御酶活性的提高有关。本研究初步揭示了印度梨形孢诱导烟草抗黑胫病的相关机理,为进一步深入研究烟草黑胫病的生物防治及其机理提供基础资料。     

叶面喷施尿素对葡萄氮代谢相关基因表达的影响

本研究利用VitisEST数据库中EST序列片段结合RT-PCR方法克隆了与氮代谢相关的硝酸盐还原酶（NR）,亚硝酸还原酶（NiR）,谷氨酰胺合成酶（GS）,谷氨酸脱氢酶（GDH）和天冬酰胺合成酶（AS）的基因,并对它们进行亚细胞定位分析。同时,采用半定量RT-PCR和荧光定量PCR法研究葡萄叶面喷施不同浓度尿素后5个基因的表达变化。结果表明,5个基因在葡萄幼叶中的表达水平显著高于老叶,说明叶面喷施尿素以喷布到幼叶对5个基因的表达水平影响显著;5个基因在不同时间段的表达水平差异不一致,但表达水平均高于对照,以0.3%和0.5%浓度的尿素对其影响明显;适当提高尿素水平既能提高氮代谢基因的表达水平,又能提高果实大小等果实指标,使其达到同步增加。喷施尿素6 h后,GS的表达量上升幅度明显高于其它基因,而NR在喷施尿素后48 h内一直保持着比较高的表达水平;NiR、AS表达量的变化趋势分别与NR、GS相一致,并且NR﹥NiR,GS﹥AS。     

胡萝卜2个DcDREB-A1类转录因子基因的克隆与比较分析

植物DREB类转录因子在植物抗逆性方面具有重要作用。本文以胡萝卜黑田五寸为材料,基于胡萝卜转录组数据,通过RT-PCR方法克隆出2个DREB-A1类转录因子基因Dc DREB-A1-1和Dc DREBA1-2。序列分析显示,这2个基因均没有内含子,长度分别为780bp和669bp,分别编码259和222个氨基酸;预测其蛋白质相对分子质量分别为29.0KD和24.71KD,p I值分别为4.32和4.63。通过对氨基酸亲水/疏水性进行分析,发现这2个转录因子属于亲水性蛋白。实时定量PCR分析表明,Dc DREB-A1-1和Dc DREB-A1-2基因在胡萝卜不同组织中的表达量不同,分别在叶和根中表达量最高。低温(4℃)、高温(38℃)、盐(0.2 mol·L-1Na Cl)、干旱(200 g·L-1PEG)不同时间段处理表达分析显示,Dc DREB-A1-1在低温、高温、盐和干旱胁迫下被显著诱导,高温、盐和干旱处理1 h后表达量达到最高,分别比对照增加14倍、7倍、7倍,低温处理下2 h表达量最高,是对照的18倍;而Dc DREB-A1-2在高温、低温和盐处理下响应明显,高温处理1 h后表达量为对照的12倍,低温和干旱处理下8 h基因表达量分别比对照增加2.4倍、6.2倍,说明2个基因在响应逆境胁迫时表达不同。胡萝卜响应非生物逆境胁迫是一个复杂的过程,本试验对深入研究胡萝卜抗逆分子机制,提高胡萝卜逆境抗性等方面具有较为重要的意义。     

接种蚯蚓对潮土氮素矿化特征的影响

在室内恒温（20℃）培养，间歇破坏性采样（于培养后第2、6、13、20、27、41、55天采样）的条件下，研究蚯蚓活动对土壤中氮素矿化特征的影响。共设置4个处理：（1）对照处理-不接种蚯蚓不添加秸秆（S）；（2）仅接种蚯蚓不添加秸秆处理（E）；（3）不接种蚯蚓仅添加秸秆处理（O）；（4）接种蚯蚓并添加秸秆处理（0E）。结果表明：在整个培养时期中，仅接种蚯蚓处理（E）的土壤铵态氮和硝态氮含量均较对照处理（S）有显著提高（P〈0．05），分别较同期对照处理（S）高出0．54-5．71倍和0．04-2．01倍；接种蚯蚓添加秸秆处理（OE）的土壤中的铵态氮含量较同期仅添加秸秆处理（0）增高了0．42-7．26倍，并达到显著性差异（P〈0．05），但两处理的硝态氮含量无显著性差异（P〉0．05）。从整体水平来看，无论是否添加秸秆，接种蚯蚓处理（E，OE）的土壤矿质氮水平均较相应的对照处理（S，O）有显著提高（P〈0．05），到培养55d时，分别增加了1．93倍和2．36倍。仅接种蚯蚓处理（E）的土壤氮素矿化速率和累积矿化速率较对照处理（S）有显著提高（P〈0．05）；添加秸秆后，无论是否接种蚯蚓，土壤累积矿化速率均为负值，表现为对土壤矿质氮的净固定，而从土壤氮素矿化速率的变化来看，前期的各个时间段中主要表现为矿质氮的净固定，后期逐渐表现为一定程度的矿化。此外，仅接种蚯蚓处理（E）的全氮含量除了培养2d外，其他培养时期均与对照处理（S）有显著差异（P〈0．05），到培养55d时，其全氮含量较同期对照处理（S）增高了6．55％。相关性分析结果表明，在仅接种蚯蚓处理（E）中，培养前后蚯蚓鲜重的减少量与土壤全氮含量之间存在极显著正相关（P〈0.01），根据一元线性方程将蚯蚓损失的鲜重换算为蚯蚓体损失的氮量后发现，在整个培养期中蚯蚓体本身排泄的氮大约占土壤全氮含量增加量的百分比为42％-100％。     

1H nuclear magnetic resonance-based metabolomic characterization of wines by grape varieties and production areas

(1)H NMR spectroscopy was used to investigate the metabolic differences in wines produced from different grape varieties and different regions. A significant separation among wines from Campbell Early, Cabernet Sauvignon, and Shiraz grapes was observed using principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). The metabolites contributing to the separation were assigned to be 2,3-butanediol, lactate, acetate, proline, succinate, malate, glycerol, tartarate, glucose, and phenolic compounds by PCA and PLS-DA loading plots. Wines produced from Cabernet Sauvignon grapes harvested in the continental areas of Australia, France, and California were also separated. PLS-DA loading plots revealed that the level of proline in Californian Cabernet Sauvignon wines was higher than that in Australian and French Cabernet Sauvignon, Australian Shiraz, and Korean Campbell Early wines, showing that the chemical composition of the grape berries varies with the variety and growing area. This study highlights the applicability of NMR-based metabolomics with multivariate statistical data sets in determining wine quality and product origin.     

Toward the authentication of wines of Nemea denomination of origin through cleaved amplified polymorphic sequence (CAPS)-based assay

In the present study, we developed a cleaved amplified polymorphic sequence (CAPS)-based assay as a first attempt to detect fraud in grapevine musts with a long-term objective to establish an analytical methodology to authenticate wines of Nemea denomination of origin (Agiorgitiko). The analytical assay makes use of a single nucleotide polymorphism that discriminates Agiorgitiko and Cabernet Sauvignon varieties. The latter grape variety is one of the major adulterants for Nemea wines. Agiorgitiko grapevine must was spiked with Cabernet Sauvignon in several ratios (v/v) from 50 down to 10%, and the subsequent mixes were subjected to alcoholic microfermentation. DNA was extracted from all mixture samples up to the end of the fermentation process and was subjected to the CAPS assay. Both standard agarose gel and lab-on-a-chip capillary electrophoresis illustrated the ability of the method to detect the presence of Cabernet Sauvignon down to 10% throughout the whole fermentation process.     

Detailed characterization of proanthocyanidins in skin, seeds, and wine of Shiraz and Cabernet Sauvignon wine grapes (Vitis vinifera)

The distribution of proanthocyanidin (PA) polymer lengths, proanthocyanidin concentration at each polymer length, and polymer composition were determined in the seed, skin, and wine of Shiraz and Cabernet Sauvignon grape berries grown in southeast Australia. PA was fractionated by semipreparative high performance liquid chromatography (HPLC) and analyzed by phloroglucinolysis and HPLC to report the degree of polymerization (DP), concentration, and composition at 11 DP values in seed and wine and 21 DP values in skin. In skin, the highest PA concentration was observed at a DP of 31 in Shiraz and 29 in Cabernet Sauvignon representing 15% of the total PA in both varieties. The distribution of seed PA had the highest concentration at a DP of 7 in Shiraz and 6 in Cabernet Sauvignon representing around 30% of the total PA. In the wine PA distribution, the highest concentration was observed at a DP of 11 in Shiraz and 9 in Cabernet Sauvignon representing around 26 and 32% of the distribution, respectively. A second peak in wine PA concentration was observed at the largest DP of 18 in Shiraz and 15 in Cabernet Sauvignon representing around 20% of the distribution. The composition in wine did not vary at different DP, but the proportion of epicatechin gallate varied in seed PA less than 4 DP. The proportion of epigallocatechin increased with increasing DP in skin PA. Wine PA had a DP range and composition similar to the distribution of skin PA between DP 4 and 18 suggesting that larger skin PAs are not extracted into wine. This study provides information that could be used to target the important PA fractions in grapes that need to be measured to understand (or predict) PA extraction into wine and eventual mouthfeel.     

Dying-arm disease in grapevines: diagnosis of infection with Eutypa lata by metabolite analysis

Dying-arm disease in grapevines, produced by infection with the ascomycete Eutypa lata, is responsible for major production losses in vineyards. Dieback of the shoots and cordon is believed to be due to acetylenic phenol metabolites produced by the fungus. To identify specific metabolites that could potentially be used for diagnosis of infection, eight E. lata isolates were grown in vitro on hot water extracts from grape varieties with various degrees of tolerance to the foliar symptoms of E. lata dieback. HPLC analysis showed that eutypinol was consistently produced in large amounts, together with smaller amounts of methyleutypinol and eulatachromene; eutypine, the putative toxin, was produced solely on Sauvignon Blanc extract and then in only barely detectable amounts. When E. lata isolates from Cabernet Sauvignon and Merlot were grown on identical media, the amounts of metabolites produced differed significantly between isolates but the pattern of metabolites was quite similar, with eutypinol again predominating. The consistent production of eutypinol indicated that this was the most suitable metabolite for which to analyze in order to diagnose the presence of E. lata. Extraction and analysis of grapevine tissues exhibiting symptoms of dieback failed to show the presence of any metabolites. However, when infected cordon sections were placed in water and cultured for 5 days, eutypinol was readily detected in the aqueous solution; metabolites were not produced from uninfected tissue. This provides a method for detection of infected tissue and indicates that the toxic metabolites react at the point of production, disrupting the vascular structure and inhibiting transport of nutrients, rather than being translocated to tissues that exhibit symptoms.     

Identification of impact odorants in Bordeaux red grape juice, in the commercial yeast used for its fermentation, and in the produced wine

The aroma extract dilution analysis method was used to detect the impact odorants of Bordeaux Cabernet Sauvignon and Merlot wines extracts, as well as those of the extracts of the corresponding Cabernet Sauvignon juice and dry yeasts used for its fermentation. The wines and the yeasts were extracted using dichloromethane, and the juice was extracted using Amberlite XAD-2. Structural identification of the impact odorants using gas chromatography-mass spectrometry and atomic emission detection (sulfur acquisition) was achieved after enrichment of these extracts by silica gel and Affi-Gel 501 chromatography. The same odorants (with the exception of dimethyl sulfide among 48) were detected in both wine extracts, with about the same flavor dilution (FD) factors. The 18 impact odorants detected in the Cabernet Sauvignon juice and dry yeast extracts were also found in the wine extracts. The odorants with the highest FD factors were 3-(methylsulfanyl)propanal, (E,Z)-nona-2, 6-dienal, and decanal in the juice extract, 2-methyl-3-sulfanylfuran, 3-(methylsulfanyl)propanal, 2-/3-methylbutanoic acids, and phenylethanal in the dry yeast extract, and 2-/3-methylbutanols, 2-phenylethanol, 2-methyl-3-sulfanylfuran, acetic acid, 3-(methylsulfanyl)propanal, 2-/3-methylbutanoic acids, beta-damascenone, 3-sulfanylhexan-1-ol, Furaneol, and homofuraneol in the wine extracts. Determination of the odor thresholds of some of these impact odorants was carried out.     

Characterization of Vitis vinifera L. Cv. Carménère grape and wine proanthocyanidins

A formal compositional study of the proanthocyanidins of Vitis vinifera L. cv. Carménère was conducted in this work. We first characterized the polymeric proanthocyanidins of Carménère skins, seeds, and wines. In addition, the wine astringency was analyzed and compared with Cabernet Sauvignon. Although Carménère wines had a higher proanthocyanidin concentration and mean degree of polymerization than Cabernet Sauvignon wines, the former wines were perceived as less astringent. The low seed/skin proportion in Carménère wines as compared to other varieties, as evidenced by the reduced number of seeds per berry and the higher amount of epigallocatechin subunits of Carménère wine proanthocyanidins, could explain this apparent paradox.     

Identification of Cabernet Sauvignon anthocyanin gut microflora metabolites

Anthocyanins are polyphenol antioxidants that have been shown to prevent many chronic diseases, including colon cancer. The compounds are largely metabolized by various enzymes and bacteria in the large intestine, and the health benefits of consuming foods rich in anthocyanins could be due mostly to the effects of these metabolites. In this study, the contents of the large intestine of pigs were used to model anthocyanin metabolism because pig and human intestinal microflora are similar. An anthocyanin extract from Cabernet Sauvignon grapes that contained delphinidin-3-glucoside, petunidin-3-glucoside, peonidin-3-glucoside, and malvidin-3-glucoside was employed. The extract was incubated anaerobically in the contents of the large intestine of freshly slaughtered pigs for 0, 0.5, and 6 h (final concentrations of 20.9, 28.2, 61.4, and 298.0 microM of the above anthocyanin compounds, respectively, at t = 0 h). Anthocyanins and their metabolites were measured by LC-ESI-MS. After 6 h, anthocyanins were no longer detected, and three metabolites were identified as 3-O-methylgallic acid, syringic acid, and 2,4,6-trihydroxybenzaldehyde. Results from this study suggest that consumption of Cabernet Sauvignon grape anthocyanins could lead to the formation of specific metabolites in the human gut, and it is possible that these metabolites offer the protective effect against colon cancer attributed to anthocyanin consumption.     

Variations in the profile and content of anthocyanins in wines made from cabernet sauvignon and hybrid grapes

To detect adulteration of wine, it has been proposed that the ratio of acetylated to p-coumaroylated conjugates of nine characteristic anthocyanins can be used to determine whether a wine is derived from Cabernet Sauvignon or hybrid grapes. If the ratio is >3, then a wine is classified as being derived from Cabernet Sauvignon grapes. This test has significant commercial implications as it is being used to decide whether Cabernet Sauvignon-labeled wines are genuine and can be imported into Germany. To assess whether this is a valid approach, 24 wines were analyzed, 4 of which were made from hybrids and 20 from Cabernet Sauvignon, with vintages ranging from 1993 to 2000. Only 13 of the Cabernet Sauvignon wines contained all nine of the "characteristic" anthocyanins, and the ratio of acetylated to p-coumaroylated derivatives varied from 1.2 to 6.5. It is evident that the use of the anthocyanin ratio method is flawed and that examination of the whole anthocyanin profile and/or investigation of the proportion of monoglucoside and acetylated anthocyanins is a better approach to distinguish between hybrid and Cabernet Sauvignon wines.     

黄瓜枯萎菌粗毒素对不同抗性黄瓜种子萌发及幼苗胁迫作用研究

利用黄瓜枯萎菌粗毒素处理不同抗性品种黄瓜,研究粗毒素对黄瓜种子萌发及幼苗生长的影响,结果表明:粗毒素对不同抗性的黄瓜胚根生长和侧根分化均有抑制作用,且随处理浓度增加,抑制作用愈加明显,但粗毒素对感病品种的抑制作用强于抗病品种。粗毒素胁迫使黄瓜胚根的细胞膜透性、MDA含量、可溶性糖含量增加,其中感病品种的细胞膜透性和MDA含量高于抗病品种,但可溶性糖含量低于抗病品种。在粗毒素处理36 h内,抗感品种黄瓜幼苗的PAL、PPO和POD酶活性均呈现先升高后降低的趋势,其中PAL和PPO的峰值出现在粗毒素处理12 h,而POD的峰值出现在粗毒素处理24 h,抗病品种的3种酶活性峰值高于感病品种。表明抗病品种对黄瓜枯萎菌粗毒素胁迫的抵抗能力强于感病品种。