荔枝胚胎发育的研究

荔枝胚胎发育的研究邱燕平,张展薇,丘荣熙（广东省农科院果树所,广州５１０６５０）（广东教育学院,广州５１０３０３）ＳＴＵＤＹＯＮＥＭＢＲＹＯＡＮＤＥＮＤＯＳＰＥＲＭＤＥＶＥＬＯＰＭＥＮＴＯＦＬＩＴＣＨＩＣＨＩＮＥＮＳＩＳＳＯＮＮ￥ＱｉｕＹａｎ－ｐｉｎ...     

荔枝胚胎发育过程的观察

荔枝是我国南方重要的亚热带果树之一,福建的栽培面积仅次于广东,列居全国第二,在果树业中占有重要位置。福建龙溪地区的主栽品种中,根据种子的发育状况,可分为三个类型:(1)正常大核型,其胚胎发育正常,成熟果实一般具有发育完好的种子,即大核。代表品种如乌叶、早红等;(2)败育型,其胚胎发育受阻,成熟果实种子败育,即所谓焦核。代表品种如绿荷包;(3)部分败育型,同一棵树或同一果穗所结的果实,其胚胎发育部分正常、部分败育。因此,成熟果实既有正常的大核果,又有一定比例的焦核果。代表品种如兰竹。     

无核荔枝胚胎发育时期蛋白质图谱分析

通过二维聚丙烯酰胺凝胶电泳(2DE)以及计算机辅助的图像分析技术,对荔枝开花后20d的正常与败育胚蛋白质图谱进行了初步分析。结果表明,正常胚总蛋白质斑点数为129,败育胚总蛋白质斑点数为130,其中24个蛋白质点在两种胚中的表达丰度没有明显变化,35个蛋白质点在表达丰度上有明显差异,55%的蛋白则发生了蛋白质缺失、增加以及位置改变等变化。这两种蛋白质组的表达差异说明了胚内蛋白质成分在其败育过程中发生了变化,这些蛋白可能参与了胚败育的调节和控制。     

荔枝胚珠中多胺含量变化与胚胎发育的关系

荔枝胚胎发育与胚珠中３种多胺（ＰＡｓ）含量及其比例变化关系密切。试验结果表明：正常发育的胚珠中腐胺（Ｐｕｔ）、亚精胺（Ｓｐｄ）和精胺（Ｓｐｍ）的含量在胚胎发育的各个阶段均高于败育胚珠,并在花后７ｄ即达到最高值,其中Ｐｕｔ的含量最高,随后都呈下降趋势。但正常胚珠中Ｓｐｍ含量在花后２２至３１ｄ（球形胚至心形胚发育阶段）均有所回升,而败育胚珠无此现象。败育胚珠中的Ｓｐｄ和Ｓｐｍ在胚胎败育期的下降速度显著     

醮核荔枝胚胎发育相关蛋白质的分离及初步鉴定

通过二维聚丙烯酰胺凝胶电泳以及计算机辅助的图像分析技术,对荔枝开花后40d的正常与败育胚蛋白质图谱进行初步分析。结果表明,100个蛋白质点在表达丰度上有明显差异;选择仅在正常发育胚胎胶上表达的蛋白点15个和仅在败育胚胎胶上表达的蛋白点50个,进行基质辅助激光解吸附电离飞行时间质谱（MALDI—TOFMASS）分析,鉴定出9个与胚发育相关的蛋白,这些蛋白可能参与了胚败育的调节和控制。     

荔枝蝽象胚胎发育研究——Ⅰ.胚胎发育过程中形态和生理的变化及温度对发育速度的影响

荔枝蝽象(Tessaratoma papillosa Drury.简称荔蝽)是我国华南地区荔枝及龙眼的重要害虫。关于荔蝽的发生规律、自然抗药性及化学防治的研究,近年来赵善欢等(1964年)已进行了一系列的研究。解放前,关于荔蝽的生物学特性,陈梦士(1934年)亦进行了观察,但直至现在,对于荔蝽的胚胎发育及不同发育期的生理指标尚缺乏详细的资料。本文的目的在于观察不同发育期荔蝽胚胎形态的特征及胆碱酯酶活性、吸氧量、脂肪含量、水份含量和卵重的变化。     

半夏胚胎发育的研究

半夏的大小孢子发育均属正常类型。只是种子成熟时没有胚的分化,而是形成一个原球茎。生长点的分化过程类似于珠芽的形成。种子发芽时原球茎突出种皮,在生长点下面的细胞,经过分裂,形成一个膨大的结构。根、叶原基的分化是在发芽过程中逐步完成的。     

乌龟胚胎发育的研究

根据乌龟胚胎发育过程中形态特征和内部结构的变化,将其胚胎发育从卵产出到幼龟孵出划分为28期。分别描述了各期的主要特征。并就乌龟发育分期的标准,原肠胚的形成,羊膜尾褶和后羊膜管,早期胚胎的自身运动等问题进行了讨论。     

海金沙胚胎发育的研究

利用石蜡切片法研究了海金沙胚胎发育过程。合子的第一次分裂面垂直于颈卵器的长轴,产生两个相等的子细胞,靠近颈卵器颈部的营养器官原始细胞和远离颈部的基足原始细胞。前者发育成子代孢子体的营养器官,后者发育成基足。胚胎在32细胞阶段后,第一叶顶端细胞与第一根顶端细胞几乎同时发生。第一叶突出帽状体之后,由第一叶基部保留下来的茎干顶端细胞产生第二叶。据营养器官的形态结构判断,在海金沙胚胎发育中最早出现的营养器官是叶和根。     

家蝇胚胎发育的研究——Ⅰ.早期胚胎发育

家蝇的胚胎发育还没有人作过观察,目前有关家蝇胚胎发育的叙述都是根据Fish(1947)对丝光绿蝇(Lucilia sericata Meigen)的胚胎发育的推测(West,1951)。家蝇目前已不仅是昆虫毒理学的试验材料,也是昆虫生理及生物化学的重要试验材料;为此,把它的胚胎发育弄清楚是一个基础工作。我们在昆虫不育性的试验中,也显示出有许多种     

荔枝核化学成分的研究

利用硅胶柱色谱、聚酰胺柱色谱、制备薄层色谱等方法,从荔枝（Litchi chinensis Sonn．）核乙醇提取物中分离出9个化合物,根据理化性质和波谱分析鉴定了7个化合物,分别为胡萝卜苷（daucosterol,1）、原儿茶醛（protocatechuic aldehyde,2）、原儿茶酸（protoeatechuic acid,3）、表儿茶素（（-）-epicafichin,4）、procyanidin A2（5）、procyanidin A1（6）、芦丁（rulin,7）。化合物2,4-7为首次从荔枝核中分得,化合物2．4、5为首次从该属植物中分离得到。     

荔枝胚蛋白质的提取方法

以不同体积的Tris-HCl(0.1mol／L,pH8．8)为提取液,结合不同含量(以胚鲜重计)的PVP40,对怀枝、黑叶和桂味等荔枝(Lithi chinensis)品种的胚蛋白质进行提取。结果表明,提取液体积为胚鲜重的5倍(ml g-1 FW),并加入15％的 PVP40时,提取蛋白质的效果最好,可用于荔枝胚可溶性蛋白质含量的测定;胚乳蛋白质的提取则以等体积的提取液(内含2％的PVP40)为佳。加入10% PVP40的胚蛋白提取液可直接进行SDS-PAGE电泳,用10倍于蛋白质提取液体积的乙醇沉淀胚和胚乳的蛋白提取液,可得到最佳的SDS-PAGE电泳效果。     

Cloning and Expression Analysis of Litchi (Litchi Chinensis Sonn.) Polyphenol Oxidase Gene and Relationship with Postharvest Pericarp Browning

Polyphenol oxidase (PPO) plays a key role in the postharvest pericarp browning of litchi fruit, but its underlying mechanism remains unclear. In this study, we cloned the litchi PPO gene (LcPPO, {"type":"entrez-nucleotide","attrs":{"text":"JF926153","term_id":"350601663","term_text":"JF926153"}}JF926153), and described its expression patterns. The LcPPO cDNA sequence was 2120 bps in length with an open reading frame (ORF) of 1800 bps. The ORF encoded a polypeptide with 599 amino acid residues, sharing high similarities with other plant PPO. The DNA sequence of the ORF contained a 215-bp intron. After carrying out quantitative RT-PCR, we proved that the LcPPO expression was tissue-specific, exhibiting the highest level in the flower and leaf. In the pericarp of newly-harvested litchi fruits, the LcPPO expression level was relatively high compared with developing fruits. Regardless of the litchi cultivar and treatment conditions, the LcPPO expression level and the PPO activity in pericarp of postharvest fruits exhibited the similar variations. When the fruits were stored at room temperature without packaging, all the pericarp browning index, PPO activity and the LcPPO expression level of litchi pericarps were reaching the highest in Nandaowuhe (the most rapid browning cultivar), but the lowest in Ziniangxi (the slowest browning cultivar) within 2 d postharvest. Preserving the fruits of Feizixiao in 0.2-μm plastic bag at room temperature would decrease the rate of pericarp water loss, delay the pericarp browning, and also cause the reduction of the pericarp PPO activity and LcPPO expression level within 3 d postharvest. In addition, postharvest storage of Feizixiao fruit stored at 4°C delayed the pericarp browning while decreasing the pericarp PPO activity and LcPPO expression level within 2 d after harvest. Thus, we concluded that the up-regulation of LcPPO expression in pericarp at early stage of postharvest storage likely enhanced the PPO activity and further accelerated the postharvest pericarp browning of litchi fruit.     

壶瓶碎米荠(Cardamine Hupingshanesis)胚和胚乳发育的研究

壶瓶碎米荠（Cardamine Hupingshanesis.）是十字花科中以营养体繁殖的多年生植物,在自然条件下,种子不易萌芽成苗。本文以碎米荠、油菜为对照材料对壶瓶碎米荠的胚胎发育进行了观察。壶瓶碎米荠胚发育经球形胚、心形胚、鱼雷形胚和子叶胚四个时期,属柳叶菜型。胚乳发育属核型发育类型,其胚胎发育过程和成熟胚结构与节状聚积和胚乳吸器,而壶瓶碎米荠和碎米荠无此类结构。通过胚胎发育观察证明,壶瓶碎米荠种胚结构完整,其种子不易萌发成苗可能与其特殊的生理原因有关。     

In Vitro Embryo Culture and Induction of Multiple Shoots in Lychee (Litchi chinensis Sonn.)

Immature embryos of different sizes and ages from commercialvarieties of lychee (Litchi chinensis Sonn.) were cultured ina range of different media. Embryos as small as 3 mm could becultured using in vitro techniques and subsequently grown intoplants. MS solid medium with 2% sucrose supplemented with 150ml l^–1 coconut water was most effective in stimulatingthe germination of immature lychee embryos. Embryos of lycheewere treated to induce adventitious buds from embryonic shootsas a means of achieving multiplication. The different varietiesexhibited differences in response, with Bengal embryonic shootsproducing 15 adventitious buds after pretreatment with 100 mgl^–1 BAP for 3 h. Root formation was achieved in 65% ofadventitious shoots using MS medium supplemented with 0.5 mgl^–1 NAA and activated charcoal. These plants were successfullydeflasked and grown on in the glasshouse. This technique providesof means of producing some multiple shoots from lychee embryosand has value for multiplication in a breeding program wherea method of micropropagation is unavailable. Litchi chinensis Sonn., lychee, embryo culture, multiple shoots, in vitro     

无核荔枝胚胎异常发育的研究

 无核荔枝是荔枝中较为珍贵的品种,有丰富的假种皮,种子多退化,属焦核品系。在胚胎发育过程中,小孢子四分体形成后,虽然出现有些小孢子形状不规则,原生质少或无等现象,但也有发育正常的花粉粒。而大孢子在各个发育阶段均会出现各种败育现象,因此无法形成胚,这是产生焦核的主要原因。淀粉粒的积累和分布对胚囊和雄配子体的发育起着重要作用,其分布具有一个动态的变化途径,这与挂绿荔枝有相同之处。     

Influence of Irradiated Pollen on Embryo and Endosperm Development in Kiwifruit

Development of seeds following pollination with irradiated pollenwas studied inActinidia deliciosa(kiwifruit) ‘Hayward’.Pollinations were carried out using two different sources ofpollen (‘Tomuri’ and ‘Matua’) irradiatedwith gamma rays at doses of 700 and 900 Gy. Non-irradiated crosseswere used as controls. Pollen irradiation had little effectonin vitropollen germination. Irradiated pollen affected seedset and seed content, and induced the formation of parthenogeneticembryos. In comparison to the control, the embryo growth ratewas slower and the endosperm contained very low amounts of storageproducts. Seed set was significantly reduced following bothdoses of irradiation. Two types of seeds were observed: (1)seeds with endosperm only; and (2) seeds with both embryo andendosperm. The proportion of seeds containing endosperm onlywas almost ten-fold higher than those containing both embryoand endosperm. Embryo production by gamma-irradiated pollenwas genotype- and dose-dependent. The induction of parthenogenesiswas higher following gamma ray doses of 900 Gy than 700 Gy,which suggests the ‘Hertwig Effect’; the best efficiencywas obtained with ‘Tomuri’ pollen. Ploidy levelof parthenogenetic embryos was evaluated by nuclear size (area)with the use of image analysis. There was a large differencein embryo nuclei size between control and parthenogenetic embryos(mean size 90.8 and 49.1 µm², respectively). It is concludedthat parthenogenetic embryos represent trihaploids.Copyright1998 Annals of Botany Company. Actinidia deliciosa, kiwifruit, pollen irradiation, induced parthenogenesis.     

水稻胚与胚乳分化发育中的内源多胺

稻胚发育过程中,其内源多胺以腐胺、亚精胺为主。在幼胚分化期,腐胺和亚精胺的含量很高;幼胚分化完成时,其含量急剧下降;直至分化后期才趋稳定。在胚及胚乳发育时期,还出现一种未知多胺X_(22),其含量除在胚分化完成时较少外,在胚发育的其他各期中,含量则一直很高。DNA和蛋白质含量的变化,从分化期开始递增直至物质积累成熟期,其趋势均相同。多胺可能参与胚与胚乳中核酸和蛋白质合成的调节。