草莓9–顺式–环氧类胡萝卜素双加氧酶基因FaNCED的克隆及表达分析

 采用RT-PCR和RACE技术从草莓果实中克隆ABA合成途径中关键基因FaNCED,该cDNA全长2 228 bp,具有完整的开放阅读框（ORF）,共1 827个碱基,编码609个氨基酸。序列分析表明,FaNCED编码的氨基酸序列与其他植物的NCED蛋白有很高的同源性。系统进化树分析显示,草莓NCED与同为蔷薇科的湖北海棠聚为一类,与橙、柠檬、温州蜜柚、葡萄等非跃变型果实NCED蛋白亲缘关系较近。实时荧光定量PCR分析发现,FaNCED基因在草莓根、茎、叶、花萼和果实中都有表达;在果实成熟过程中FaNCED的表达出现两次高峰,分别在大白果期和红果期,且在红果期表达量最高,并与ABA积累相吻合;FaNCED在果实采后1 d表达略有下降,之后急剧上升,ABA含量变化与FaNCED基因的表达基本一致。FaNCED可能参与调控ABA的合成并在草莓成熟中起一定作用。     

草莓果实发育过程中糖、pH值及ABA水平变化趋势

为了揭示草莓果实发育和成熟的生理机制,本研究以杜克拉草莓7个时期的果实为试材,分析果实发育过程中pH值、可溶性糖和ABA变化趋势.结果表明,果实发育前期可溶性糖积累以葡萄糖和果糖为主,蔗糖很低;果实进入始熟和成熟,蔗糖分别有一次快速积累的过程,全红时蔗糖含量是果糖和葡萄糖的1.25倍.以纯白期pH值最低点为界,pH值先...     

番茄成熟突变体果实贮藏过程中细胞超微结构的变化

 对含有rin基因的番茄成熟突变体02612果实在贮藏过程中细胞超微结构的变化进行了研究。结果表明, rin突变体果实在贮藏期间果肉细胞壁完整、叶绿体、线粒体结构保持完好。rin突变体的耐贮性与其果肉细胞在贮藏过程中能够保持完整的结构有关。     

钾肥处理对草莓果实糖积累及相关酶活性的影响

以以色列品种“S3”草莓为试材,研究了不同钾肥处理对草莓蔗糖、果糖、葡萄糖含量及相关代谢酶的影响。结果表明:钾肥的施用可以促进草莓果实蔗糖、果糖、葡萄糖含量的增加,蔗糖是构成草莓可溶性糖的主要成分,果糖和葡萄糖含量基本相当,随着钾肥施用量的增大,施钾量为360kg·hm-2时,可溶性糖含量最高;在花后42d蔗糖合成酶(SS)活性最高,适量钾肥显著提高了草莓发育后期蔗糖合成酶的活性,其活性在草莓果实发育中后期与蔗糖、葡萄糖积累呈显著正相关,在果实发育前期与果糖积累呈负相关;蔗糖磷酸合成酶(SPS)在蔗糖合成积累中起着重要的作用,草莓成熟过程中,SPS活性总体呈上升趋势,与蔗糖含量呈极显著正相关;在草莓果实成熟过程中酸性转化酶(AI)、中性转化酶(NI)活性变化较大,总体在果实成熟过程中呈先下降后上升的趋势,且与果实中蔗糖含量呈负相关。钾离子对蔗糖代谢的相关酶具有重要的调节作用,适量的钾肥处理提高了蔗糖合成酶(SS)、蔗糖磷酸合成酶(SPS)、酸性转化酶(AI)、中性转化酶(NI)活性,从而提高生长期果实中蔗糖、果糖和葡萄糖积累量。     

草莓果实成熟软化相关基因研究进展

对草莓果实细胞壁水解酶和伸展蛋白在成熟软化时的功能和作用进行了阐述,并对已报道的相关编码酶（或结构蛋 白）的基因进行了描述和评价。指出纤维素酶、果胶裂解酶和伸展蛋白是参与草莓果实成熟软化的关键因子;并提出利用RNA 干扰等生物技术对关键基因进行调控,降低相关酶的mRNA表达,提高草莓果实硬度的研究思路。     

磷脂酶D在果实发育和成熟过程中的作用

 结合作者的研究工作着重综述了磷脂酶D催化的磷脂代谢途径与特性、磷脂酶D与果实发育和成熟的关系、磷脂酶D参与的衰老相关的生理生化进程和磷脂酶D基因、蛋白质结构及其表达模式, 并探讨了磷脂酶D在果实发育和成熟进程中的可能作用机理。     

Fruit yield and quality of strawberry plants transformed with a fruit specific strawberry pectate lyase gene

Two transgenic strawberry lines (Pel 1 and Pel 3) containing the open reading frame of a fruit specific strawberry pectate lyase gene (FaplC) under the control of the CaMV35S promoter have been obtained to evaluate the role of this gene on fruit softening. Ripen fruits from both lines showed a significant down-regulation of FaplC, being the percentage of silencing of 84 and 71% on Pel 1 and Pel 3, respectively. The agronomic behaviour of transgenic plants was evaluated during two consecutive years. Fruit set increased in the two transgenic lines when compared with control plants, although Pel 1 showed a significant reduction on fruit weight. Firmness of full ripen fruits from Pel lines was significantly higher than control fruits, while color and soluble solids were not affected. The increase of firmness in Pel lines was maintained when ripe fruits were stored for 3 days at 25 °C. Histological analysis of ripe fruits showed lower intercellular spaces and a higher degree of cell to cell contact area in transgenic fruits when compared with controls. Altogether, these results suggest a direct relationship between pectate lyase gene expression and strawberry fruit softening.     

Physiological changes during development and ripening of ‘Perlita’ muskmelon fruits

‘Perlita’ muskmelon fruit (Cucumis melo L.) grown under both field and greenhouse conditions were sampled at 10-day intervals for 50 days after pollination to determine changes in pH, soluble sugars, total protein, β-carotene, tissue firmness, cell wall composition, cell wall degrading enzymes, total calcium and bound calcium. The sucrose content of fruits increased substantially after 40 days post-anthesis, which compensated for the significant decreases in glucose and fructose content. Acidity and total protein remained relatively unchanged throughout the sampling period. β-carotene increased from 0.3% (w/w) at 10 days post-pollination to 2.7% (w/w) in 50-day old melons. Tissue firmness measured by shear force decreased by 400% between 30 and 50 days postanthesis. No significant changes in cellulose, hemicellulose and pectin content of the cell wall were detected. Cx-cellulase and pectinmethylesterase activity were detected but remained constant or declined during ripening; polygalacturonase activity was not detected. The major compositional changes in developing and ripening fruit were sucrose, glucose, fructose and β-carotene. In spite of dramatic decreases in tissue firmness, concomitant changes in crude cell wall fractions and associated cell wall degrading enzymes were not detected.     

Firmness,pectin components and cell wall hydrolases of mango fruit following low-temperature stress

Summary

Mango (Mangifera indica L. cv. Nam Dokmai) fruits chilled for 21 d at 48C were firmer than non-chilled fruits during subsequent ripening at 258C. The cell wall contents of chilled fruit contained less water-soluble pectin, more ammonium oxalate-soluble pectin and less alkali-soluble pectin than non-chilled fruits. The increase in ammonium oxalate-soluble pectin was at the expense of the alkali-soluble pectin and correlated with high pectinesterase activity in the fruit from low-temperature storage. Polygalacturonase and b-galactosidase activities were inhibited in chilled fruits. The relationships between delay in loss of firmness, and differences between activity of cell wall enzymes and pectin content of chilled and non-chilled fruits are discussed.     

草莓果实ABA积累关键酶基因FaNCED1和FaBG1转录水平的分析

为探讨草莓果实发育过程中ABA积累的酶学调控机制,文章以花后1～4周的‘杜克拉’草莓7个时期果肉为试材,利用实时荧光定量PCR分析ABA积累关键酶基因FaNCED1和FaBG1转录水平。结果表明,在整个草莓果实发育过程中,果肉中FaNCED1基因表达水平明显地分为两个持续上升阶段,即从小绿期至白熟期和始红期至全红期,但二者之间还存在一个短暂下降期,即白熟期至始红期;果肉中FaBG1基因表达水平呈逐渐降低的趋势。结合果肉中ABA含量分析发现,FaNCED1基因转录水平呈现与ABA水平相似的变化趋势。本研究主要得出以下结论:(1)退绿和着色是草莓果实发育成熟过程中两个突出事件;(2)果肉中ABA水平两次持续快速积累分别与草莓果实退绿和着色相偶联;(3)果肉中ABA含量主要由FaNCED1基因决定,而与FaBG1基因关系不大。以上结果为从分子水平调控草莓果实中ABA含量,进而调控果实的发育奠定研究基础。     

Role of surface morphology in determining the ripening behaviour of tomato (Lycopersicon esculentum Mill.) fruits

Surface characteristics of fruit mainly determine the diffusibility of gases across its boundary and also the extent to which moisture being lost from the fruit. The present study has been carried out with an objective to find out the association between the ripening related processes and surface morphology of tomato (Lycopersicon esculentum Mill.) fruit. Tomato fruits of two contrasting varieties; Pusa Gaurav (a slow ripening type) and Pusa Ruby (a relatively fast ripening type) were evaluated for physiological loss in weight (PLW), rate of respiration, effect of hypoxia on ripening, dimension of stem scar region and distribution of lenticels along with hair base cell of trichomes on the surface of the fruits. Results showed that fruits of Pusa Gaurav showed significantly lower PLW, rate of respiration, surface area of stem scar region, density of lenticels along with hair base cell of trichomes and slower ripening under hypoxia condition in comparison to Pusa Ruby. More resistance towards gaseous exchange and therefore lower O₂ to CO₂ ratio inside the fruits of Pusa Gaurav in contrast to Pusa Ruby in view of the differences in the surface morphological features in these two varieties could possibly be assigned as one of the reasons responsible for the obtained varietal difference in ripening and related changes.     

草莓FaMDHAR和FaGR的克隆与表达分析

为了探讨草莓中单脱氢抗坏血酸还原酶（Monodehydroascorbate reductase,MDHAR）和谷胱甘肽还原酶（Glutathione reductase,GR）基因的功能,采用同源克隆的方法从‘丰香’草莓中克隆得到cDNA全长序列,并采用实时定量PCR方法对其在不同组织和不同发育阶段果实中的表达模式进行分析。结果表明：草莓MDHAR基因cDNA（FaMDHAR,GenBank登录号：KP025946）全长1 305 bp,编码434个氨基酸,分子量约为47 kD,含有保守的FAD结合结构域,定位于细胞质中。GR基因cDNA（FaGR,GenBank登录号：JQ339738）开放阅读框全长1 491 bp,编码496个氨基酸,分子量为53 kD,定位于细胞质中。FaMDHAR在各组织中均有表达,在成熟果实中表达量最高,叶和花次之,根中最低;在果实发育过程中FaMDHAR在绿果期有相对较高的表达,随后急剧增加,到白熟期最高,之后下降并维持在相对稳定水平。FaGR在叶和花中表达较高,在成熟果实中较低;在果实发育过程中,表达量从小绿期呈现增加趋势,至转红果期达最高,随后逐渐下降,在成熟果实中较低。果实发育过程中酶活性变化呈现出与各自基因表达量相似的变化规律。经过4 ℃低温处理24 h后的草莓叶片中,FaMDHAR相对表达量较对照显著增加,而FaGR无显著变化。草莓中FaMDHAR和FaGR表达存在时空差异,并对低温逆境响应存在差异。     

反义乙烯受体FaEtr2基因对草莓的转化

通过根癌农杆菌EHA105介导转化草莓叶片,在卡那霉素选择压下连续分化选择、扩繁和生根培养,获得了26株转乙烯受体FaEtr2反义基因植株。PCR和Southern blot检测证明,该基因已经整合草莓染色体中。Northern分析表明,转入的反义基因对靶基因都有部分的抑制。对果实乙烯释放量测定表明,转基因草莓中乙烯释放量降低。     

Relationship between the oil and sugar content in olive oil fruits from Moraiolo and Leccino cultivars during ripening

In our previous work (Cherubini et al., 2009), sugar concentration was proposed as an accurate, reproducible index for technological ripening of olive oil fruits. During 2009 crop season, sugar and oil content of olive oil fruits from Moraiolo and Leccino cultivars were measured during ripening. The sugar content of olive oil fruits was determined both by titration and by using a portable refractometer.     

草莓果实磷脂酶D活性的测定

以草莓果实为试材,建立了适合草莓果实组织磷脂酶D制备及活性测定的技术体系。以放射标记的磷脂酰胆碱为底物,在1mL反应体系中,膜蛋白的含量以5-10μg为宜;25℃反应温度下,反应时间以5-10min为宜;反应体系的适宜pH为6.5-7。对草莓果实不同组分磷脂酶D酶学特性的进一步分析认为,该技术体系的建立有助于进一步探讨磷脂酶D在草莓果实发育成熟和衰老进程中的作用。     

梨果实发育中Ca2+在果肉细胞的定位及变化研究*

 用焦锑酸钾沉淀的细胞化学方法, 研究了􀀂 幸水 梨果实发育中果肉细胞的焦锑酸Ca²⁺ 定位变化及其与细胞超微结构的关系。结果表明: ( 1) 在未受精之前, 果肉细胞内未检测到Ca²⁺ 沉淀颗粒, 细胞核内的染色质少且染色淡, 细胞质的细胞器数量也少; ( 2) 受精后果肉细胞呈现大量的Ca²⁺沉淀颗粒, 主要分布在细胞核、细胞质、质体以及叶绿体外膜上, 含Ca²⁺沉淀颗粒的质体非常膨大, 导管和初期发育的石细胞内也密集分布Ca2+ 沉淀颗粒; ( 3) 受精1 周后果肉细胞的Ca²⁺移向细胞之间的连接处; ( 4) 生理落果的细胞和导管中Ca²⁺没有或极少, 但有的细胞内沿液泡膜有Ca²⁺分布; ( 5) 受精3~ 4 周后, 果肉细胞中很难检测到Ca²⁺沉淀颗粒, 此状态一直持续到果实采收, 但果实腐烂前Ca²⁺沉淀颗粒沿果肉细胞 壁两侧出现。就Ca2+ 在果实发育中的作用及与细胞超微结构的关系等进行了讨论。     

草莓果实软化机理及调控研究进展

综述了草莓果实软化过程中细胞及呼吸作用的变化；细胞壁水解酶（多聚半乳糖醛酸酶、果胶甲酯酶、纤维素酶）在果实成熟软化过程中的活性及其作用；内源激素（乙烯、生长素、赤霉素、细胞分裂素、脱落酸）在果实成熟软化过程中的含量变化及其与果实软化的关系；钙对草莓果实成熟的影响及其与软化的关系。总结了近年来控制果实软化的主要措施，包括草莓果实的采前处理和采后处理，介绍了草莓果实的采前保护、采前喷钙、采后冷藏、气调贮藏、涂膜处理、辐射电磁处理、热处理以及防腐保鲜剂等技术的应用情况。     

几种果实不同组织总RNA提取及质量分析

以菊水梨为试验材料,通过2种改良CTAB法对果实不同成熟阶段提取的总RNA质量和产量的变化的影响,研究果实不同成熟阶段果皮和果肉总RNA提取的质量和产量,并针对梨果实不同成熟阶段采用不同的改良CTAB法,以较低的成本从果实中提取完整性好、质量高的总RNA。同时提取近成熟的红富士葡萄、富士苹果和丰香草莓3种果实总RNA。总RNA的质量和产量分析结果表明,方法Ⅰ和Ⅱ分别适用于成熟度较低和较高梨果实总RNA的提取,方法Ⅰ也适用于其它3种近成熟果实总RNA提取。并通过RT-PCR验证,所提取的总RNA可以满足基因克隆和表达等分子生物学实验的要求。     

The effect of preharvest calcium sprays on quality attributes,physicochemical aspects of cell wall components and susceptibility to brown rot of peach fruits (Prunus persica L. cv. Andross)

This study was planned to evaluate the effect of preharvest calcium sprays on physicochemical aspects of cell wall components, activity patterns of pectin-modifying enzymes and susceptibility to brown rot (Monilinia fructicola) of peach fruits (Prunus persica L. cv. ‘Andross’). Six or 10 preharvest calcium sprays were applied with two different formulas (calcium chloride and an ethylenediamine tetraacetic acid (EDTA) chelated calcium form) in equimolar calcium concentration (0.12% Ca, w/v). After harvest, peach fruits were cold stored (0 °C, 95% R.H.) up to 4 weeks. Calcium content increased significantly in the calcium-sprayed peaches both in the peel (25–42%) and in the flesh (11–17%), 1 d after harvest. Calcium chloride sprays were more effective than those of chelated calcium, whereas the increase of calcium content in the fruits seems to be unaffected by the 6 or 10 calcium sprays. The increase of cell wall calcium corresponded to increase of calcium in the insoluble pectin fraction, whereas no differences were detected in the calcium of water-soluble pectin fraction. Ethylene production, respiration rate, uronic acid content and activity of pectin-modifying enzymes did not indicate substantial differentiations by preharvest calcium sprays as ripening progressed after harvest or cold storage. Calcium sprays resulted in significant decrease of brown rot development, although no effect on disease incidence was recorded.     

草莓果实发育过程中糖代谢相关基因的表达分析

 为了分析草莓（Fragaria × ananassa Duch.）果实发育过程中可溶性糖积累与相关基因表达量的关系,以‘杜克拉’草莓7个发育时期的果实为试材,利用实时荧光定量RT-PCR的方法,分析果实发育中蔗糖转运蛋白及糖代谢关键酶（酸性转化酶、蔗糖合成酶和蔗糖磷酸合成酶）4个基因的表达量以及可溶性糖含量的变化趋势。结果表明,蔗糖、葡萄糖和果糖含量随着果实发育都呈逐渐增加的趋势,尤其蔗糖积累与果实成熟的关系较密切。随着果实发育,蔗糖磷酸合成酶基因表达量呈逐渐增加的趋势,尤其在果实发育的后期增加较快;蔗糖合成酶和酸性转化酶基因表达量除了白熟期和转色期外,呈逐渐降低的趋势;蔗糖转运蛋白基因表达量呈先升后降,降后又升的变化趋势,尤其在白熟期前急剧增加。结合可溶性糖和基因变化趋势分析表明,蔗糖转运蛋白和蔗糖磷酸合成酶在草莓果实发育过程中发挥着重要的作用。