Genomic characterization of self-incompatibility ribonucleases (S-RNases) in loquat (Eriobotrya japonica Lindl.) (Rosaceae, Pyrinae)

Loquat (Eriobotrya japonica Lindl.) is a fruit tree species of the Pyrinae subtribe of the Rosaceae that behaves as self incompatible. Since self-incompatibility in the Rosaceae is of the gametophytic type where a stylar ribonuclease (S-RNase) controls the female function of pollen–pistil recognition, consensus primers derived from the alignment of S-RNase sequences from other Pyrinae species were used to search for S-RNases in loquat. As a result, the first four S-RNases were sequenced for this species. The genomic sequences obtained showed the structural features of Pyrinae S-RNases. Moreover, microscopic observations of pollen tube growth in the style confirmed the inter-(in)compatibility relationships predicted from the molecular analyses. Phylogenetic analysis of the deduced amino acid sequences with other Pyrinae S-RNases confirmed that divergence of S-alleles in loquat and the Pyrinae predated speciation. This study reports for the first time the genomic characterization of S-RNases in loquat, providing a sound basis for an appropriate selection of pollinator cultivars and an adequate design of breeding programs.     

枇杷果皮响应高温强光胁迫的蛋白质组分析

为探讨枇杷[Eriobotrya japonica(Thunb.)Lindl.]果皮在高温强光胁迫下的蛋白质组分变化,采用蛋白质组学方法分析了果实日灼抗性差的枇杷种质‘WDYDB’果皮蛋白质对高温强光胁迫的应答反应。结果表明,在自然高温强光胁迫与遮光处理(对照)下,枇杷果皮蛋白质双向电泳图谱中表达量差异在2倍以上的蛋白点共有31个;通过MALDI-TOF-TOF/MS质谱分析成功鉴定出26个差异蛋白点,包括11个下调蛋白和15个上调蛋白。根据这些蛋白功能,可将其分为防御应答、碳水化合物和能量代谢、光合作用、其它等4类蛋白。同时,对这些蛋白质在高温强光胁迫下的功能和作用进行了讨论。这些差异蛋白质参与了枇杷对高温强光胁迫的响应。     

枇杷果实采后生理与保鲜技术研究进展(综述)

本文综述枇杷果实采后生理与保鲜技术的研究现状,并提出枇杷保鲜技术发展方向。     

枇杷叶尖焦枯病叶脉组织电镜观察初报

用超薄切片和电镜观察技术,对枇杷叶尖焦枯病进行组织病理学观察。结果发现,病株叶脉韧皮部的筛管及伴胞内有大量丝状菌体状物存在,其大小为37~100×1500~?nm,横切面形态为近圆形、椭圆形或不规则短杆状,纵切面形态为菌丝状,而对照的无病实生苗木中则无。据此初步认为,丝状菌体状物可能与枇杷叶尖焦枯病的发生有关。     

19个枇杷杂交新品种(系)的SSR鉴定和指纹图谱构建

为探究枇杷(Eriobotryajaponica)杂交品种真实性和DNA指纹图谱,对新育成的19个枇杷杂交品种(系)进行SSR标记鉴定分析。结果表明,从已发表的89对SSR引物中筛选出扩增条带清晰稳定的多态性引物19对,在24份枇杷材料中共扩增到83条带,每对引物平均扩增4.37条,PIC值为0.234～0.983,平均为0.764。经12对具有父本特征带多态性引物鉴定, 19个杂交新品种(系)全部为真杂种,真杂种率为100%。UPGMA聚类分析表明,19个杂交新品种(系)的遗传相似系数为0.728～0.969,与杂交亲本‘新白2号’和‘贵妃’聚为同一个大类,并可细分为4个亚类,红肉与白肉的枇杷品种(系)间无明显划分。同时利用8对多态性SSR引物组合,构建了19个枇杷杂交新品种(系)的分子指纹图谱。这为枇杷品种鉴定、新品种权保护和杂交育种提供重要参考依据。     

响应面法优化微波辅助提取枇杷叶中黄酮类化合物

为充分利用枇杷叶,研究微波辅助乙醇浸提法从枇杷叶中提取黄酮类化合物的工艺。首先研究了影响提取效果的因素:不同体积分数的乙醇溶液、料液比、时间及微波功率。在单因素试验基础上利用响应面分析法,确定了各因素与总黄酮得率之间的关系,并以总黄酮得率为响应值做响应面和等高线图。研究表明,枇杷叶中黄酮类化合物提取最佳工艺为乙醇体积分数50%,料液比1∶30,提取时间3 min,微波功率500 W,最佳条件下总黄酮得率为2.04%,与理论推导值2.07%相比,误差为1.4%,黄酮类化合物响应方程的回归分析表明此方法合理可行。本研究对枇杷叶总黄酮提取优化的研究为进一步探讨枇杷叶的深度开发提供了理论参考依据,并为枇杷叶总黄酮的生物活性研究提供材料。     

果实成熟度和贮藏条件对枇杷种子无菌萌发的影响

为探讨影响枇杷(Eriobotrya japonica Lindl.)种子萌发的因素,将种子播种于1/2MS培养基80 d,研究了成熟度和贮藏条件对枇杷种子萌发的影响。结果表明,‘软条白沙’、‘宁海白’、‘塘科白沙’和‘大红袍’转色期和完熟期采收的种子的萌发率和苗高无显著差异;而‘大房’完熟期采收种子的萌发率较高。‘洛阳青’枇杷果实或种子经4℃贮藏5 d和10 d,萌发率和苗高与未贮藏种子的无显著差异,而贮藏期达20 d或更长时,贮藏种子的萌发率和苗高显著下降。在不同温度下贮藏的枇杷种子萌发率和苗高存在差异,‘早钟6号’和‘太平白’枇杷在15℃和4℃的优于25℃的,但适宜贮藏时间分别不超过15 d和30 d。因此,在转色期采收果实以及适度贮藏果实或种子可以保持枇杷种子的萌发能力。     

不同贮藏温度对枇杷果肉木质化及相关酶活性的影响

将“解放钟”枇杷(Eriobotrya japonicaL ind l.cv.Jiefangzhong)果实于4℃和12℃下贮藏,研究了不同贮藏温度对采后枇杷果肉木质化及相关酶活性的影响。结果表明,4℃贮藏的枇杷果肉苯丙氨酸解氨酶(phenylalan ineammon ia lyase,PAL)、4-香豆酸辅酶A连接酶(4-coum arate coenzym e A ligase,4-CL)、肉桂醇脱氢酶(c innamyl alco-hol dehydrogenase,CAD)和过氧化物酶(peroxidase,POD)活性高于12℃贮果,而多酚氧化酶(polyphenol oxidase,PPO)活性低于12℃贮果。4℃贮藏的枇杷果肉硬度和木质素含量不断增加,果肉木质化较为严重;12℃贮藏的枇杷果肉硬度和木质素含量低于4℃贮果,果肉木质化程度较低。低温诱导PAL和POD活性的上升是导致枇杷果肉发生木质化的主要原因。     

Effect of detergents, trypsin and unsaturated fatty acids on latent loquat fruit polyphenol oxidase: basis for the enzyme's activity regulation

The effects of detergents, trypsin and fatty acids on structural and functional properties of a pure loquat fruit latent polyphenol oxidase have been studied in relation to its regulation. Anionic detergents activated PPO at pH 6.0 below critical micelle concentration (cmc), but inhibited at pH 4.5 well above cmc. This behavior is due to a detergent-induced pH profile alkaline shift, accompanied by changes of intrinsic fluorescence of the protein. Gel filtration experiments demonstrate the formation of PPO-SDS mixed micelles. Partial PPO proteolysis suggest that latent PPO losses an SDS micelle-interacting region but conserves an SDS monomer-interacting site. Unsaturated fatty acids inhibit PPO at pH 4.5, the strongest being linolenic acid while the weakest was gamma-linolenic acid for both, the native and the trypsin-treated PPO. Down-regulation of PPO activity by anionic amphiphiles is discussed based on both, the pH profile shift induced upon anionic amphiphile binding and the PPO interaction with negatively charged membranes.