猕猴桃软化过程中阶段性专一酶活性变化的研究

猕猴桃（Ａｃｔｉｎｉｄｉａ ｄｅｌｉｃｉｏｓａ Ｃ．Ｆ．Ｌｉａｎｇ ｅｔＡ．Ｒ．Ｆｅｒｇｕｓｏｎ． ｃｖ． Ｑｉｎｍ ｅｉ）果实采后的软化过程表现为两个明显的阶段,第一阶段软化较快,此时对软化起主要作用的阶段性专一酶是淀粉酶;第二阶段软化速度变慢,此时起主要作用的阶段性专一酶是多聚半乳糖醛酸酶和纤维素酶。乙烯形成酶（ＥＦＥ）的活性高峰出现在两个软化阶段之间,它所引起的乙烯释放对软化有促进作用,因此ＥＦＥ也是与果实软化有关的阶段性专一酶。但是,果胶甲酯酶（ＰＭＥ）的活性变化与果实的软化无相关关系,过氧化物酶（ＰＯＸ）、过氧化氢酶（ＣＡＴ）和ＳＯＤ的活性高峰出现在果实完全软化以后,因此不是果实软化的阶段性专一酶     

SDP型猕猴桃保鲜剂研制及其应用研究

本文介绍,以菜油磷脂和其它天然产物为原料,研制出SDP型猕猴桃保鲜剂,应用于“海沃特”果实的保鲜贮藏。在产地简易通风库中,常温贮藏3～5月,好果率>70％,比对照多出好果8～18％,Vc含量高于对照9～37.3％,果实外观、口感、风味均正常,此外,初步探讨了保鲜机理,提出了天然营养性防腐保鲜的新概念。     

采收期和贮藏温度对金艳猕猴桃品质的影响

研究了金艳猕猴桃(Actinidia chinensis ×A. eriantha) 4个不同采收期(Ⅰ~Ⅳ)在常温[(23±1)℃]贮藏60 d,以及低温(4℃)贮藏2、4、6月后常温货架14 d内果实品质和果肉色彩的变化。结果表明：采收期Ⅰ和Ⅳ的果实在常温贮藏过程中呼吸高峰出现早、失重快,可滴定酸(TA)、维生素C(Vc)含量和果肉硬度迅速下降;而采收期Ⅱ和Ⅲ的果实呼吸高峰出现晚,保持了较高的TA、Vc含量和果肉硬度。4个采收期猕猴桃果实在常温贮藏期间,果实亮度L*、色度C*和色彩角h显著降低;而在低温贮藏后常温货架期间,果实色彩角h相对于亮度L*和色度C*变化更加明显。研究表明,采收期Ⅱ和Ⅲ为金艳猕猴桃的适宜采收期,色彩角h可以作为适宜的指标反映猕猴桃果实的后熟与衰老,而低温贮藏6个月金艳猕猴桃仍能保持较好的食用品质和色彩。     

一种高效提取猕猴桃DNA和RNA的方法

在总核酸提取方法(PS法)的基础上,经多次实践改进,得出一种以高盐低pH的HAc-NaAc缓冲体系提取总核酸的简便方法,可以从富含多糖、多酚时猕猴桃叶片和花蕾中提取同时含有DNA和RNA的总核酸.所得的总核酸在LiCl溶液中选择性沉淀RNA,从而有效地分离出DNA和RNA样品.紫外分光光度法和琼脂糖凝胶电泳分析表明,所提取的DNA和RNA具有较高的纯度和完整性.通过样品DNA的PCR和样品RNA的RT-PCR,认为所提取的DNA样品和RNA样品能够满足分子生物学试验的基本要求.     

猕猴桃POD基因的克隆和表达分析

为了解猕猴桃POD基因的表达调控功能,采用RT-PCR技术从‘米良1号’猕猴桃(Actinidiadeliciosa‘Miliang-1’)克隆了2个POD家族成员基因(AdPOD27和AdPOD64)。结果表明,AdPOD27和AdPOD64开放阅读框分别为984和957 bp,预测分别编码327和318个氨基酸,GenBank登录号分别为MF774100和MF774101。AdPOD27和AdPOD64为亲水性碱性蛋白,属于植物亚铁红素依赖Ⅲ型POD超家族成员,含有信号肽、跨膜螺旋结构和磷酸化位点,亚细胞定位预测分别定位于线粒体和细胞外。q RT-PCR结果表明,Ad POD27在脱落酸(ABA)和4℃处理时表达量急剧上升,而AdPOD64只在ABA处理时表达量显著提高。此外,AdPOD27表达量与POD活性、AdPOD64表达量均存在显著相关性。因此,AdPOD27和AdPOD64可能在猕猴桃果实软化、低温响应和ABA诱导等过程中发挥重要的调控功能。     

Process of infection of armored scale insects (Diaspididae) by an entomopathogenic Cosmospora sp

Several species in the fungal genus Cosmospora (synonym Nectria) (anamorph Fusarium) are specialist entomopathogens of armored scale insects (Diaspididae), known to cause periodic epizootics in host populations. Inconsistent mortality rates recorded under laboratory conditions prompted a study into the process of infection of armored scale insects by this fungus. Scale insect mortality following exposure to a Cosmospora sp. (Culture Collection Number: CC89) from New Zealand was related to insect age, with reproductively mature insects having a significantly higher infection rate than immature insects. Examination using scanning electron microscopy found no evidence that the fungus penetrated directly through the wax test (cap) of the scale insect or through the un-lifted interface between the test and the substrate on which the insect resided. However, fungal hyphae were observed growing beneath the test when the test of the reproductively mature insect lifted away from the substrate for the purpose of releasing crawlers, the mobile pre-settled juveniles. Once the hyphae of CC89 advanced under the test, germ-tubes readily penetrated the insect body through a number of natural openings (e.g. spiracles, vulva, stylet), with mycosis observed within seven days after inoculation. Direct penetration through the cuticle of the scale insect was not observed.     

Expression of soybean b-1,3-endoglucanase cDNA and effect on disease tolerance in kiwifruit plants

Kiwifruit was transformed with a soybean β-1,3-endoglucanase (EC 3.2.1.39) cDNA under the control of the cauliflower mosaic virus (CaMV) 35S RNA promoter. The introduced gene was expressed in young leaves of the transformants. Assays of protein extracts from young leaves showed an increase in enzyme activity in many transformants, the transformant with the highest level of enzyme activity having an about sixfold increase over the control plants. When leaves from control and three transformants were inoculated with Botrytis cinerea, which causes gray mold disease, the disease lesion areas for two transformants were smaller than on control plants. Received: 5 March 1998 / Revision received: 19 October 1998 / Accepted: 27 October 1998     

猕猴桃叶DNA的AFLP分析

以猕猴桃幼叶为材料提取基因组DNA并进行AFLP扩增。对主要实验步聚包括DNA纯化、DNA的酶切、酶切片段与接头的连接、PCR扩增、电泳、以及银染等方面的反应参数进行比较和优选,初步摸索出适合于猕猴桃叶为材料的AFLP程序,并得到较为清晰可辩的AFLP指纹图谱,为开展猕猴桃遗传多态性研究和在分子水平上开展物种生物学分析提供一个实用的手段。     

秦美猕猴桃叶片最佳再生系统的建立

利用正交等试验,系统开展了秦美猕猴桃叶片再生系统建立研究,确定了秦美猕猴桃叶片不定芽诱导最佳培养基及激素配比为MS＋6－BA 5mg/L NAA 0.1mg/L,不定芽诱导生根最佳培养基及激素配比为1／2MS＋NAA 0．01mg/L IBA0.5mg/L GA 1mg/L,该实验结果为通过叶盘法开展农杆菌介导的猕猴桃遗传转化奠定了良好基础。     

Involvement of the ubiquitin/proteasome pathway in the organisation and polarised growth of kiwifruit pollen tubes

We recently reported the involvement of the ubiquitin pathway in microgametophyte development, and a direct role for the 26S proteasome in regulating pollen tube emergence in kiwifruit. Here we show that the ubiquitin/proteasome proteolytic pathway is involved not only in early kiwifruit pollen tube organisation, but also in maintaining polarised growth of tubes. By immunofluorescence analysis we show that ubiquitin and ubiquitin-protein conjugates are distributed mainly at the apex of emerging tubes, in both untreated pollen grains and pollen grains treated with MG132, an inhibitor of proteasome function. In the latter case, polysiphonous germination occurred and all the emerging areas were highly fluorescent. By adding MG132 to pollen when normal tube growth had already been established, accumulation of ubiquitin-protein conjugates, as well as a drastic reduction in tube growth and dramatic modifications of tube tip morphology were observed. Significantly, differential interference contrast microscopy analysis demonstrated that the clear zone was largely reduced or absent, and the nuclei were disconnected in their movements, reaching, in some cases, the extreme apex of the tip. These findings provide evidence that the ubiquitin- and proteasome-dependent proteolytic system could modulate the abundance and/or activity of key regulatory proteins involved in pollen tube emergence and polarised growth.