利用CODEHOP设计简并引物克隆镰刀菌果胶酶基因片段

利用CODEHOP(Consensus-degenerate hybrid oligonucleotide primers)设计真菌果胶酶基因片段的简并引物,并对设计的多对引物进行筛选,比较了普通PCR和Touchdown-PCR(TD-PCR)程序的扩增效果,并对产物进行了测序、比对和分析。结果表明:利用CODEHOP设计简并引物可信性强,阳性率高,能够从供试菌株中获得与目的片段大小相近的产物。利用TD-PCR程序扩增比普通PCR扩增效果好。扩增产物序列BLASTX比对和分析结果表明,产物片段编码的氨基酸序列与镰刀菌属来源的果胶酶氨基酸片段相似性均超过90%,说明所扩增的序列即为镰刀菌果胶酶基因片段。     

Study of Defense-related Gene Expression in Grapevine Leaves and Berries Infected with Botrytis cinerea

Defense responses of grapevine towards Botrytis cinerea were investigated. The expression of genes coding for proteins involved in defense were studied: (a) phenylalanine ammonia-lyase (PAL) and stilbene synthase (StSy), (b) an acidic chitinase (VCH3) and a basic chitinase (VCHIT1b), and (c) a polygalacturonase inhibitor protein (PGIP). Since no PGIP was known in grapevine, a complete cDNA sequence was first characterized by PCR and RACE-PCR amplifications. RNAs isolated from infected leaves and infected berries were analysed by semi-quantitative and real-time RT-PCRs. In infected leaves, the expression of PAL, StSy, PGIP and VCH3 genes occurred 6hours post inoculation (hpi). Increase of VCHIT1b gene expression was delayed (24hpi). Maximum levels of induction of these genes were observed at 48hpi, except for the VCH3 gene (24hpi). Activation of these defense responses was not sufficient to stop B. cinerea spread. In berries, no VCH3 gene expression was detected. Maximum levels of induction were observed in stage 3 (loss of berry colour and abundant production of conidia) for the PAL and PGIP genes, and in stage 4 (shrivelled berry) for the StSy and VCHIT1b genes.     

青枯菌胞外酶对木麻黄的致病作用研究

对三个致病性不同的木麻黄青枯菌ＰｓｅｕｄｏｍｏｎａｓｓｏｌａｎａｃｅａｒｕｍＥ．Ｆ．Ｓｍｉｔｈ产生的胞外酶活性测定显示,聚半乳糖醛酸酶（果胶酶）活性在各菌株之间无明显差别,但纤维素酶活性差别明显,并随菌株的致病性上升而增加,菌株致病性与酶活性具有高度正相关（ｒ＝０．９６）。     

半乳糖醛酸酯酶的活性对芒果硬度的影响

以新摘大台农芒果为原料,对其采后采取不同贮藏处理方式,对贮藏期间芒果的硬度、多聚半乳糖醛酸酶(PG)的活性和果胶含量的变化进行了研究。结果表明,在贮藏过程中,PG活性提高,导致原果胶水解为可溶性果胶,原果胶平均含量从贮藏初期的1.721%下降到最后的0.9982%,而可溶性果胶从0.1076%上升到0.9452%,总果胶含量也稍微呈上升趋势,从1.8342%上升到1.8969%;PG酶活性先上升至最高点,然后稍微有所下降,从11.34个酶单位增大到17.10个酶单位后又回落到16.03个酶单位,芒果硬度从0.234kg/cm下降为0.162kg/cm,果22实发生了软化。以不经过任何处理的芒果为空白对照,比较了5种不同涂膜保鲜方法的效果,并且从这些保鲜方法中,筛选出较好的保鲜方法是涂130mg/kg甲基托不津+1.3%壳聚糖+90%甘油,贮藏期可延长7d。     

不同耐贮性梨贮藏中果胶分解酶活性变化比较

以南果梨、延边小香水梨和苹果梨为试材,比较分析耐贮性不同的梨果实贮藏中果胶分解酶活性变化.结果表明,耐贮性强的苹果梨贮藏中PG酶活性很低,贮藏20 d出现活性高峰;而β-半乳糖苷酶活性高,但没有明显的活性高峰.耐贮性弱的南果梨和延边小香水梨果实贮藏中PG酶活性很高,但没有明显的活性高峰;南果梨的β-半乳糖苷酶活性高于苹果梨,而延边小香水梨的则低于苹果梨,但南果梨和延边小香水梨分别于贮藏15 d和10 d时出现明显的活性高峰.说明梨的耐贮性与贮藏中PG酶活性高低有关,但与活性高峰无关;耐贮性强弱与β-半乳糖苷酶活性大小无关,而与活性高峰出现与否有关,且耐贮性越弱,活性高峰出现越早.     

Polygalacturonase-inhibiting protein activity in cantaloupe fruit as a function of fruit maturation and tissue origin

Netted cantaloupe (Cucumis melo var. cantalupensis cv. Magnum 45) were harvested from 5 to 35 days postanthesis. The fruit of each age group were divided into exocarp, outer mesocarp, mid mesocarp, inner mesocarp, placenta, and seed. Each tissue was extracted and assayed for polygalacturonase-inhibiting protein (PGIP) activity against polygalacturonases (PGs) from three fungal pathogens of cantaloupe fruit. The PGIP activity of all tissues except placenta was high from the flower stage through the first week of fruit development but decreased markedly between 5 and 10 days postanthesis. PGIP activity against Phomopsis cucurbitae PG remained high and nearly constant in placental tissue throughout fruit development. However in this same tissue, PGIP activity against Fusarium solani PG decreased during fruit development to about 25% of its level in the 5-day-old fruit. This differential change in PGIP activity toward the two PGs suggests that different forms of the inhibitor are expressed between early and late stages of cantaloupe fruit development. The results also illustrate the importance of using multiple pathogen enzyme systems that can provide an opportunity for more accurate elucidation of mechanisms involved in the host–pathogen interaction. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture. All programs and services of the US Department of Agriculture are offered on a nondiscriminatory basis without regard to race, color, national origin, religion, sex, age, marital status, or handicap. The article cited was prepared by a USDA employee as part of his/her official duties. Copyright protection under US copyright law is not available for such works. Accordingly, there is no copyright to transfer. The fact that the private publication in which the article appears is itself copyrighted does not affect the material of the US Government, which can be freely reproduced by the public.     

乌龙茶做青过程中果胶酶活性及相关生化成分的动态变化

为了探明乌龙茶做青过程中果胶酶活性及相关生化成分的变化规律，选用金萱、白叶单枞的新梢，设置了新工艺摇青和在新工艺基础上加大强度摇青2个处理，并以在制品的不摇青处理作为新工艺的对照，研究了各处理过程在制品的果胶酶(PG)活性动态变化，同时，研究了Ca^2 处理对PG活性的影响，并结合相关生化成分的变化探讨了PG在做青过程中对乌龙茶品质形成的生化效应。结果表明，茶鲜叶晒青后，PG活性明显上升，并在摇青期间的一摇或二摇后PG活性达到峰值；在新工艺基础上加大强度摇青，PG活性逐步下降，摇青处理的PG活性明显高于不摇青处理；Ca^2 能显著抑制PG活性，PG活性变化与可溶性糖含量消长有密切联系。     

In vitro Effect of the Phytoalexin Glyceollin I on Polygalacturonase and Oxalic Acid Production ofSclerotinia sclerotiorum

The polygalacturonase (PG) activity in culture filtrates fromSclerotinia sclerotiorum is reduced when glyceollin I, the major soybean phytoalexin, is present in the culture medium. When the enzyme activities in the culture filtrates are expressed per unit of fungal growth, PG activity decreases with increasing concentration of glyceollin I in the culture medium. The phytoalexin does not influence the isoenzyme pattern. This suggests that glyceollin I may inhibit quantitative but not qualitative enzyme production. Only the highest glyceollin I concentration tested inhibits oxalic acid production. The inhibitory effect on mycelial growth is confirmed. The data suggest a further hypothesis about the role of phytoalexin during pathogenesis.     

人工接种CGMMV后与西瓜倒瓤相关部分生理指标的研究

 本研究以辽宁省田间主栽西瓜品种“京欣”为试材,通过测定多聚半乳糖醛酸酶(PG)的活性,以及乙烯、丙酮酸及丙酮酸代谢产物—乳酸、乙醛、乙醇含量的变化,明确了西瓜感染黄瓜绿斑驳花叶病毒后上述生理指标的改变与西瓜倒瓤的关系。研究结果表明:接种处理的西瓜果实授粉后7 d和28 d PG出现2次活性高峰,PG活性过高将导致果肉软化;乙烯含量亦在授粉后21~28 d明显高于健康对照,这一变化将导致果肉水渍化败坏;无论接种早晚,西瓜果实内丙酮酸及丙酮酸代谢产物均有不同程度的积累。定植期接种处理的西瓜,授粉后21 d时西瓜果实内丙酮酸含量为健康对照的2.51倍,35 d时达健康对照4.09倍;授粉后7 d乳酸含量已显著高于对照,35 d时接种处理的西瓜果实内的乳酸含量迅速增加至43.5 μmol·g^-1FW,达健康对照的4.7倍;乙醛含量随着西瓜成熟逐渐增加,尤其是授粉后21~35 d乙醛含量增加速度极为迅速,35 d时达健康对照的26.4倍;乙醇含量授粉后7 d最高,达健康对照的10.1倍,随后有所下降。于压蔓期接种处理的西瓜果实,授粉后21 d,丙酮酸、乳酸、乙醛含量明显高于健康对照,但均低于定植期接种处理的,乙醇含量与健康对照无明显差异,也就是说,感染该病毒越早,对西瓜的生长影响越明显。