荔枝若干重要果实品质性状的QTL分析

以‘马贵荔＇ב焦核三月红＇76株F_1代群体为试材,检测了酒石酸、苹果酸、蔗糖含量和单果重4个果实性状分离的情况。结果表明,4个性状表现为连续分布,具有数量性状的典型特征,与4个果实性状连锁的QTL位点23个,其中控制酒石酸的QTL为2个,控制苹果酸的QTL为4个,控制蔗糖的QTL为12个和控制单果重的QTL为5个。各QTL的LOD值在3.15～5.61之间,可解释13.85%～88.3%的表型变异。     

荔枝果皮过氧化物酶的纯化及部分酶学性质研究

经硫酸铵分级盐析、DEAE-Sepharose和Sephadex G-75柱层析分离,从荔枝果皮中分离提纯了过氧化物酶(POD),该酶被纯化了12．5倍,产率为1．9％。经SDS-PAGE确定为单一条带。该酶最适反应温度为35℃,对热具有较强的稳定性,经75℃处理30min,酶活性只损失50％。最适pH约为6．5,但在pH4．0—8．0范围内活力仍比较稳定。该酶在25℃和0．05mol／L磷酸缓冲液(pH7．0)条件下对愈创木酚、邻苯二酚和没食子酸的Km分别是2．75、12．4和12．8mmol／L。二硫苏糖醇和抗坏血酸能完全抑制POD活性,L-半胱氨酸、柠檬酸、FeS04、GSH、SDS和ZnS04对POD活性有一定的抑制作用,而FeCl,和CuSOt对POD则有较好的激活作用。     

Somatic embryogenesis and plant regeneration of litchi (Litchi chinensis Sonn.) from leaves of mature phase trees

Embryogenic cultures were induced from leaflets from new vegetative flushes of mature ‘Brewster’ litchi trees on B5 medium containing 400 mg l⁻¹ glutamine, 200 mg l⁻¹ casein hydrolysate, 30 g l⁻¹ sucrose, 4.52 μM 2,4-D, 9.30 μM kinetin and 3 g l⁻¹ gellan gum in darkness. Embryogenic cultures consisting of proembryonic cells and masses were maintained either on semi-solid MS medium supplemented with 4.52 μM 2,4-D and 0.91 μM zeatin or as embryogenic suspension cultures in liquid medium of the same composition. Maturation of somatic embryos occurred on semi-solid MS medium with 5–20% (v/v) filter-sterilized coconut water in darkness. Recovery of plants from somatic embryos was improved with 14.4 μM GA₃ on half-strength MS medium with 0.2 g l⁻¹ activated charcoal under a 16 h photoperiod provided by cool white fluorescent lights (60–80 μmol s⁻¹ m⁻²). Plants have been successfully acclimatized in the greenhouse.     

Biology and control of the fruit borer, Conopomorpha sinensis Bradley on litchi （Litchi chinensis Sonn.） in northern Thailand

Fruit and leaf shoot of litchi （Litchi chinensis） infestation by naturally occurring populations of the fruit bprer, Conopomorpha sinensis larvae and C. sinensis parasitization rates were determined in litchi orchards in northern Thailand at high （ca. 1 400 m ASL） and low （ca. 800 m ASL） elevations with different, non-overlapping fruiting seasons. In addition, spray applications of Bacillus thuringiensis var. aizawai, the natural compound spinosad and the chemical pesticide imidacloprid were conducted in the field to test the effects of these agents on C. sinensis fruit infestation rate. Fruit infestation rate and fruit growth, studied in a low elevation orchard, were sigmoidal and showed a highly significant positive correlation （P ≤ 0.01）. Leaf shoot infestation rate in the low elevation orchard decreased in the course of the fruiting season （March until May）, but increased in the high elevation orchard, where no fruits were present within the same period of time. Together, these results indicate that females of C. sinensis clearly prefer fruits over shoots for oviposition. If no fruits are available, which was the case in the high elevation orchard during that time, they are constrained to lay their eggs on shoots. The parasitization rate of C. sinensis larvae in untreated fruits was 54.3%. The parasitoid species were Chelonus chailini （90.8%） and Phanerotoma sp. （9.2%）. In the leaf shoots, C. sinensis was found to be parasitized by Phanerotoma sp. and Apanteles briareus. The applications of the different active agents had no significant effect on C. sinensis fruit infestation compared to the untreated control. Besides the cryptic life of the larvae, this is most probably explained by the fact that C. sinensis has several generations during the fruiting season. Because of the high parasitization rates recorded, an alternative and more effective control measure than the application of pesticides might be the augmentative release of parasitoids.     

应用RAPD标记研究野生荔枝种质资源

应用RAPD方法对海南60份野生荔枝进行基因组多态性分析,20个随机引物共产生165条RAPD带,DNA片段大小在200～1500bp之间,其中121条为多态性带,占总带数的73．3％,并利用遗传距离进行聚类分析。结果表明,60份野生荔枝可归为6类,表明种群存在一定的遗传变异,也为分析野生荔枝群体间及群体内的遗传多样性探索了有效方法。     

大核和焦核"桂味"荔枝果实发育及其发育期间果皮中内源激素含量的变化比较

同一株大核和焦核"桂味"荔枝的果实生长型均呈"单S型",其果实大小差异主要由种子大小不同引起.果实发育期间,两者果皮中内源激素变化的规律大体上一致,大核果皮中GA3、IAA含量和(IAA ZRs GA3)/ABA比值均高于焦核果皮,但ZRs含量和ZRs/ABA比值则比其低,ABA含量的差异无规律可循.     

桂味荔枝花器官的发生和发育过程研究

利用SV11立体显微镜和JSM-6360LV型扫描电镜观察‘桂味’荔枝花器官的发生和发育过程。结果表明:花序原基最先发生,然后形成数个大小不等的单花原基;4个萼片原基的发生不同步,其中一侧对位先发生;6～10枚雄蕊原基以轮状方式几乎同时发生;心皮原基最后发生,2～3枚(稀4枚)心皮原基同时出现,随后进行侧向生长,逐渐合拢形成子房。雌花中,花柱、柱头分化明显,雄蕊退化。雄花中,花丝细长,花药饱满,雌蕊退化或发育不完全。两性花中,雌雄蕊发育完全。花粉粒近球形,具3孔沟,表面为条纹状纹饰。     

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。     

Identification of polysaccharides from pericarp tissues of litchi (Litchi chinensis Sonn.) fruit in relation to their antioxidant activities

A large number of polysaccharides are present in the pericarp tissues of harvested litchi fruits. A DEAE Sepharose fast-flow anion-exchange column and a Sephadex G-50 gel-permeation column were used to isolate and purify the major polysaccharides from litchi fruit pericarp tissues. Antioxidant activities of these major polysaccharide components were also evaluated. An aqueous extract of the polysaccharides from litchi fruit pericarp tissues was chromatographed on a DEAE anion-exchange column to yield two fractions. The largest amount of the polysaccharide fraction was subjected to further purification by gel filtration on Sephadex G-50. The purified product was a neutral polysaccharide, with a molecular weight of 14 kDa, comprised mainly of 65.6% mannose, 33.0% galactose and 1.4% arabinose. Analysis by Smith degradation indicated that there were 8.7% of (1-->2)-glycosidic linkages, 83.3% of (1-->3)-glycosidic linkages and 8.0% of (1-->6)-glycosidic linkages in the polysaccharide. Furthermore, different polysaccharide fractions extracted and purified from litchi fruit pericarp tissues exhibited strong antioxidant activities. Among these fractions, the purified polysaccharide had the highest antioxidant activity and should be explored as a novel potential antioxidant.     

基于SSR标记的荔枝种质遗传多样性分析

从53对本课题组自主开发设计的SSR特异引物对中筛选出22对多态性引物,对46份荔枝材料的基因组DNA进行扩增,共得到54个等位基因,其中每对引物的平均等位基因数为2.4,共获得23个特异性标记,各标记的平均观察杂合度值、平均期望杂合度和PIC值分别为0.451、0.355和0.507。其中L it6位点各数值最高,是最理想的选择标记。利用22对多态性引物对46份荔枝种质进行聚类分析,构建树状图,结果表明:大多数种质相似系数在0.63～0.95之间,说明它们之间的亲缘关系较近,并发现淮枝(广东)与淮枝(海南)两者可能为同名异物品种。     

Multiple shoot regeneration of kenaf (Hibiscus cannabinus L.) from a shoot apex culture system

 In this research, a medium was developed that would stimulate multiple shoot initiation from shoot apex explants of Hibiscus cannabinus L. (kenaf). Adventitious shoot formation on a shoot induction media supplemented with combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) (0, 0.5, 2.3 μmol·l^–1) and thidiazuron (N-phenyl-N′-1,2,3-thiadiazol-5-ylurea; TDZ) (0, 1, 5, 20 μmol·l^–1) was evaluated. Multiple shoot induction medium with 1 μmol·TDZ l^–1 resulted in the highest number of regenerated shoots per explant for all three kenaf cultivars tested (Tainung 1, Tainung 2, and Everglades 71). The 2,4-D did not enhance multiple shoot formation. Additionally, kenaf cultivars 7N and SF459 also produced multiple shoots on the induction medium with 1 μmol·TDZ l^–1. Multiple shoot clumps formed after 2 weeks in culture without callus formation. Shoots elongated and rooted within 2 weeks after subculture on a plant growth regulator-free medium. A histological study demonstrated the de novo regeneration of shoots from the shoot apex. Received: 2 February 2000 / Revision received: 30 March 2000 / Accepted: 22 June 2000     

荔枝DNA提取及RAPD扩增条件优化

为应用RAPD技术开展对荔枝种质资源的分析,以S43(GTCGCCGTCA)为引物,通过试验设计,分别研究了退火温度、模板浓度、引物浓度、dNTP浓度、Taq DNA聚合酶用量对荔枝RAPD-PCR反应的影响。建立并优化了适宜荔枝RAPD分析的扩增体系:20μL的反应体系,30ng的模板DNA度,0.25μmol/LRAPD引物、1.0UTaqDNA聚合酶,0.2μmol/LdNTP为荔枝适宜的RAPD-PCR扩增条件。     

霜疫霉菌侵染对荔枝氧化作用的影响

淮枝、桂味和糯米糍 3种荔枝经接种霜疫霉菌后 ,果皮超氧化物歧化酶 (SOD)和过氧化氢酶 (CAT)活性下降 ;而过氧化氢 (H2 O2 )含量升高 ,超氧阴离子自由基 (O2 .)产生速率增加 ,丙二醛 (MDA)积累增多 ,与果实感病指数增加相一致 ,表明霜疫霉菌侵染加速荔枝氧化作用的进程。实验还表明不同荔枝品种对霜疫霉菌抵抗力与其自由基清除能力有关     

荔枝SSR标记的研究

以无核荔枝A4号为实验材料,应用选择性扩增微卫星（SAM）法分离、克隆了100个简单序列重复（SSR）序列,其中88个非重复,可用。加上搜索数据库所获得的1个SSR序列,一共89个序列用于特异引物的设计。仅从71个序列的82个基因座设计出特异引物。合成41条特异引物(与5′锚定简并引物配对,个别相互配对),对其中的39个基因座进行检测。其中15对引物扩增出相应大小的片段,另外11对引物扩增出非预期片段。最后,以37个荔枝种质的基因组DNA为模板,从26对出带的引物中,筛选出多态性引物21对,获得了22个荔枝基因座特异性SSR标记。     

荔枝果皮总蛋白质提取及双向电泳体系的建立

用TCA-丙酮、丙酮和酚3种方法提取荔枝(Litchi chinensis Sonn.)果皮的总蛋白质,比较了蛋白产量、单向SDS-PAGE和双向电泳等方面的差异,并对双向电泳体系进行探索.结果表明:酚抽提法最佳,提取的总蛋白得率最高,蛋白在单向SDS-PAGE中形成条带数目最多,最清晰;经双向电泳分离用银染显色,可检...     

In vitro induction of multiple shoots and plant regeneration from shoot tips of mung bean (Vigna radiata (L.) Wilczek)

Conditions for plant regeneration from excised shoot tips of Vigna radiata were studied. Complete plants were regenerated directly without an intervening callus phase from shoot tips on basal medium (MS salts+B₅vitamins). Regeneration frequency varied with genotype, explant size and growth regulator combinations in the medium. Addition of cytokinins induced a variable amount of callus at the base of the shoot tip, followed by multiple shoot formation. Benzyladenine (BA), kinetin and zeatin at 5×10^-6 M each induced multiple shoots in 100% of the explants but the highest number of regenerants per explant (9) was produced with BA. The efficacy of BA for shoot multiplication was not improved when it was supplemented with naphthaleneacetic acid (NAA) or indoleacetic acid (IAA). NAA or adenine sulphate, when applied alone, induced complete plantlets. The growth regulator requirement of explants for the induction of multiple shoots varied with explant size. The shoot tip explants maintained proliferation ability on subculture. None of the treatments was effective in inducing shoot bud differentiation from callus. Regenerated shoots were rooted on MS basal medium and MS supplemented with either IAA or indolebutyric acid. The rooted plants were transferred to the field; 60% subsequently survived and grew.Abbreviations BM basal medium [MS (Murashige & Skoog 1962) salts+B₅ (Gamborg et al. 1968) vitamins] - BA 6-benzyladenine - AdS adenine sulphate - IAA indole-3-acetic acid - NAA-1 naphthaleneacetic acid - IBA indolebutyric acid     

纯氧对荔枝果实贮藏期间果皮褐变和细胞超微结构的影响

研究了纯氧对荔枝果皮褐变和细胞超微结构的影响。结果表明,100％O2显著抑制了荔枝果皮褐变,在100％O2下贮藏6d的荔枝果实,中果皮细胞的细胞器和膜系统的完整性保持较好,而细胞膜透性较低,因此,认为高氧抑制荔枝果皮褐变与其维持细胞膜的完整性相联系。     

Development and characterization of SSR markers in lychee (Litchi chinensis)

Sixteen microsatellite loci were isolated from lychee (Litchi chinensis), all of which exhibited polymorphism (two or three alleles per locus), with levels of heterozygosity ranging from 0.021 to 0.900. These loci can help assess the genetic structure of lychee.     

Intergeneric Hybridisation between Litchi (Litchi chinensis Sonn.) and Longan (Dimocarpus longan Lour.)

The breeding barriers between commercial litchi (Litchi chinensisSonn.) and longan (Dimocarpus longan Lour.) cultivars were investigatedby conducting reciprocal pollinations. This work has shown thatit is possible to generate intergeneric hybrids using litchias the female parent. Investigation of comparative in vivo pollentube growth demonstrated that there is discrimination againstcross- compared to self-pollen at all sites in the pistil. Pollentubes were frequently observed in the ovary after cross-pollinationin litchi but rarely in longan. Fruit production was reducedafter crossing in both longan and litchi. Isozyme analysis usingphosphoglucose isomerase revealed that hybrid progeny only developedwhen litchi was the maternal parent. Morphologically the hybridplants were similar to the maternal parent but leaves were smaller.Three types of seeds developed in litchi following pollinationwith longan pollen. These were (1) normal seeds with a developedtesta and embryo, (2) seeds with aborted embryos but normaltesta development, and (3) seedless fruit where the ovule remainedthe same size as at anthesis without further development ofembryo or testa. The potential germplasm available to improvethese crops within the Sapindaceae is discussed.Copyright 1994,1999 Academic Press Litchi, Litchi Longan, Dimocarpus, hybridisation, isozyme     

荔枝-旋扭山绿豆复合种植系统对荔枝害虫的生态调控作用

对营建荔枝 旋扭山绿豆复合种植系统和荔枝单一种植系统的昆虫群落结构及动态进行系统调查.结果表明,复合种植园中荔枝害虫总数显著减少,仅为单一种植园的61.27%,在果实生长关键期的5月份的数量仅是单一种植园的50.45%.旋扭山绿豆与荔枝树存在相互协调关系：两者具较少共同害虫,其害虫却存在多种共同天敌;旋扭山绿豆冬季开花为天敌昆虫越冬提供庇护场所和替代食物,两者花期基本衔接,利于害虫天敌的生存及繁殖.在荔枝开花和荔果生长期的3～6月份,复合园荔枝树上的天敌数量与害虫数量之比分别为单一园的4.22、2.34、2.28和20.63倍,有效地发挥了对荔枝害虫的控制作用.