高等植物合子离体培养研究进展

合子是高等植物个体发育的第一个细胞,在研究植物个体发育机理中有着不可替代的位置。植物体内合子细胞分裂并以胚胎发生方式形成新个体的特征在植物生物工程应用中独具特色。随着离体受精技术的发展和合子分离技术的改进,已从多种植物中得到了人工合子和从体内直接分离出了合子。合子离体培养系统的建立为研究高等植物个体发育最初阶段的合子激活机理提供了条件,也为探索利用合子胚胎发生的特征进行植物基因工程打下了基础。另外,探索异种植物间人工杂种合子的培养也是进行远缘杂交的新尝试。     

甜樱桃胚胎离体培养

甜樱桃（Cerasusavium）果实采收后贮藏在40℃下30～40d。除去果肉,将内果皮撬开后,取出胚放在无菌水中浸泡30min,70％乙醇中30s,再用0．2％HgCl2或2％～4％NaOCI表面灭菌20min,最后用无菌水冲洗4次。将胚切成两部分,一部分带有胚珠（大小为整个胚的2／3）,另一部分为子叶。将带有胚珠的外植体接种在诱导培养基（MS＋0．5ms·＊－‘6－BA＋2％蔗糖十0．8％琼脂）上,20d后观察统计胚胎发育类型。然后,将外植体发育成的植株切除或保留主根转入增殖培养基（MS＋0．5mg·L－‘6－BA＋2％蔗糖十0．8％琼脂）。切除主根的试管苗培…     

葡萄霜霉病离体接种方法的研究

用葡萄霜霉菌孢子囊悬浮液接种于葡萄叶盘,一般接种后３－４天,叶盘上可见孢子囊,接菌后５－６天,侵染率可达９５－１００％。     

棉属种间杂交胚珠的离体培养

棉属(Gossypium)的种间杂交是育种的重要手段之一。通过种间杂交,可将棉属中某些种,尤其是野生种和半野生种的优良特性引入生产上栽培的种,从而创造人们所需要的品质好、抗逆性强的新类型或新品型。这是长期以来育种工作者的愿望。同时,棉属的种间杂交对创造雄性不育系,探索棉属的分类与进化以及对于遗传育种理论的研究都有重要意义。然而,棉属的种间杂交也和其它一些植物一样存在着杂交亲本的不亲和性和杂种后代的不育性。根据胚胎学     

未成熟杏胚离体培养

ImmatureEmbryoCultureofApricotinvitroWANGYuZhu,SHIHons（PomologyandForestInstitute,AgricultureandForestryScienceAcademyofBeijing,Beijing100093）1植物名称杏（Prunusarmeniaca）极早熟品种“骆驼黄”。2材料类别盛花后41、46和57d的未成熟胚。3培养条件培养基为Tukey、SH、Norstog［KNO3160mg／L（单位下同）,Ca（NO3a）2·4H2O290,NaH2PO4·H2O800,Na2SO4200,MgSO4·7H2O730,KCl140,FeC6H5O7（1％）10,H3BO30.5,CuSO4·5H2O0．25,MnSO4·4H2O3,ZnSO4·7H2O0．5,Na2MoO4·2H2O0．…     

广藿香不同外植体离体培养的研究

以广藿香叶片、带节茎、不带节茎及根尖为材料进行离体培养,对影响离体再生的因素进行了研究。结果表明:BA有利于广藿香外植体出芽,浓度以0.1～0.5mg/L效果较好;不同外植体的出芽能力有较大差异,其中以叶片和带节茎出芽能力较强,出芽率均达100%;外植体在培养基上的接种方式对出芽也有一定影响,带节茎以形态学下端垂直插入,可以缩短出芽时间及增加单个外植体出芽的数量。无根苗生根以MT+IBA0.2mg/L培养基为好,苗的生长较为健壮。     

提高西瓜离体培养植株再生效率的研究

本文以"京欣1号"母本和"伊选"西瓜4天苗龄子叶为外植体,研究离体培养植株高频率再生体系.结果表明"伊选"子叶远轴端外植体的再生频率仅为10%,子叶近轴端外植体在5mg/LBA+0.1mg/L IAA的激素组合下植株再生频率为100%,平均每个外植体的丛生芽数在所有组合中最多,为10.3个;"京欣1号"母本子叶近轴端外植体在2mg/L BA+0.5mg/L IAA激素组合下植株再生频率为100%,平均每个外植体的丛生芽数在所有组合中最多,达6.9个.本试验条件下,子叶近轴端外植体接种4天即分化出不定芽,至再生苗的移栽仅需40天,在MS+0.1mg/L NAA的生根培养基上的生根率为97.3%,移栽成活率达98.5%.     

石蒜与萱草远缘杂交育种研究

目的：为培育石蒜属植物新品种（系）提供参考依据。方法：以石蒜属（Lycoris）、朱顶红属（Hippeastrum）、萱草属（Hemerocallis）植物为试材,采用花粉离体培养、田间杂交、胚拯救等方法,研究了石蒜属植物在远缘杂交过程中的生殖隔离。结果：1、在花粉活力测定中,所得最适宜朱顶红花粉萌发的培养基组成为：70 g/L蔗糖+5 mg/L硼酸+0.01 g/L氯化钙+50 g/LPEG-4000;萱草组花粉的最适培养基组成为：100 g/L蔗糖+0.3 mg/L硼酸+0.4 mg/L氯化钙。2、以石蒜属植物作母本,设计8个杂交组合试验,结果表明子房膨大率随杂交组合的不同而呈现差异性,其中与四倍体萱草杂交组合所得平均子房膨大率为59.81%,高于二倍体萱草组（33.12%）。3、胚拯救初期,各组合获得胚珠0～44个,其中鹿葱×萱草‘金娃娃’组合最多,中国石蒜×萱草‘金娃娃’组合最少。胚珠膨大率随杂交组合的不同出现差异,7个杂交组合的胚珠均发生膨大,膨大率在1.79%～96.15%之间。其中鹿葱×’Custard Candy’组合胚珠膨大率较高（96.15%）,稻草石蒜×’Cust...     

中国野生毛葡萄酒的香味成分分析

毛葡萄酒的香味成分由醇、酯、醛、酮等构成,3-甲基-1-丁醇是最主要的香气成分,分别占总量的34.1％（83-4-67）、34.12％（泰-12）、41.78％（商-24）;其单宁、总酚含量高,颜色深浓。毛葡萄种内3个系酒的香味成分总量、种类和各类成分的含量均存在明显差异。毛葡萄酒具有青草、烟熏、肥皂、焦糖及酸涩等香味特征。     

广西野生毛葡萄雌雄株花粉形态学研究

通过电镜扫描,对原产于广西石山地区的野生毛葡萄雌、雄株花粉形态进行比较研究。结果表明,野生毛葡萄花粉粒为小型花粉粒,雌、雄株花粉粒形状及纹饰、萌发孔特征均有较大差异:雄株花粉粒萌发孔为三孔沟类型,长球形,花粉粒P/E=1.20(P极轴值,E赤道轴值);雌株花粉粒无萌发孔,近球形,花粉粒P/E=1.12。     

中国野生毛葡萄钙调蛋白基因克隆及序列分析

于葡萄黑痘病发病盛期,用病叶压片法对高抗黑痘病的中国野生毛葡萄'商-24'接种黑痘病病原菌,采用mRNA差异显示技术进行抗黑痘病基因表达差异的研究.结果显示:(1)获得了T11GG/B0304-400、T11CC/S428-350、T11GG/S424-700、T11AG/S432-350、T11GG/S433-250、T11GG/S433-300、T11AG/S432-300、T11GG/S438-353和T11AG/S424-300等9个基因表达差异cDNA片段.其中T11GG/S438-353 mRNA片段表达在接种后3 d被诱导显著降低,并在之后2 d几乎检测不到.(2)采用RACE技术克隆了T11GG/S438-353 mRNA片段的cDNA全长序列;序列分析表明,该cDNA包含一个607 bp完整的开放阅读框架,编码149个氨基酸;其编码氨基酸序列与拟南芥、欧洲葡萄、柳杉、党参、无梗花栎、欧洲栗及寄生草钙调蛋白的一致性分别为99%、97%、94%、91%、90%、88%和77%.(3)本实验克隆到了负向调控中国野生毛葡萄抗黑痘病的钙调蛋白基因,并命名为VqCaM,其GenBank登录号为EU694099;实时荧光定量PCR结果再次验证VqCaM表达受葡萄黑痘病侵染下调,     

In vitro culture of ovaries of a viviparous gall midge

Summary Ovaries of the viviparous pedogenetic gall midgeHeteropeza pygmaea can be cultured in hemolymph obtained from X-ray-sterilized larvae of the same species. In this culture medium, formation of follicles is essentially the same as in vivo, and sometimes female larvae develop from these follicles. The ovaries of such larvae, in their turn, have been cultured in vitro to produce larvae. In this way, in vitro development from oogonium to larva has been maintained for several generations. When using hemolymph obtained from larvae grown under different conditions, the in vitro cultured ovaries produce a second type of egg which probably is male-determined. Ovarian development in vitro has been studied with differential interference contrast optics and time-lapse cinemicrography. This work was supported by the Swiss National Science Foundation Grant No. 3.2010.73.     

In vitro culture of Arabidopsis embryos within their ovules

Embryogenesis of flowering plants establishes a basic body plan with apical-basal, radial and bilateral patterns from the single-celled zygote. Arabidopsis embryogenesis exhibits a nearly invariant cell division pattern and therefore is an ideal system for studies of early plant development. However, plant embryos are difficult to access for experimental manipulation, as they develop deeply inside maternal tissues. Here we present a method for the culture of zygotic Arabidopsis embryos in vitro. The technique omits excision of the embryo by culturing the entire ovule, thus greatly facilitating the time and effort involved. It enables external manipulation of embryo development and culture from the earliest developmental stages up to maturity. Administration of various chemical treatments as well as the use of different molecular markers is demonstrated together with standard techniques for visualizing gene expression and protein localization in in vitro cultivated embryos. The presented set of techniques allows for so far unavailable molecular physiology approaches in the study of early plant development.     

In vitro propagation of cauliflower,Brassica oleracea var. botrytis for hybrid seed production

Methods for obtaining heterotic F₁ and maintaining purebred lines for breeding of Brassica oleracea are limited by absence of male sterile lines and occurrence of inbreeding depression, respectively. The use of vegetative (stem, petiole, leaf, leaf rib) and floral (peduncle, pedicel, flower bud, curd) explants of cauliflower to regenerate purebred lines for crossing were examined. Of four growth regulator treatments and explant types used, best results were obtained with curd explants on MS medium with 6-benzyladenine (cytokinin) and gibberellic acid. Although 6-benzyladenine alone promoted formation of shoots in floral explants, both 6-benzyladenine and α-napthaleneacetic acid were required for vegetative explants. Use of α-napthaleneacetic acid, however, often increased callus formation. These culture techniques to maintain purebred regenerated plants will complement newly-derived nuclear-based male sterile lines obtained by the introduction of antisense copies of the gene BcpI, which is required for pollen fertility. This revised version was published online in June 2006 with corrections to the Cover Date.     

In vitro culture of cells and tissues from the mosquito,Anopheles quadrimaculatus say

Summary Tissues and cells from various life stages and ages ofAnopheles quadrimaculatus were cultured in several media. Pupal tissue maintained in Schneider's (1) medium was the most successful system. These cells remained viable for 210 days. Xix morphologically distinguishable cell types were observed: spherical cells, two type of neuronlike cells, musclelike cells, epithelial-like cells, and polymorphic cells.     

凤梨科植物的离体培养（综述）

本文主要从快速繁殖,育种探索和种质保存三个方面介绍凤梨离体培养研究的进展,并简要介绍其他凤梨科植物离体培养的研究概况,在此基础上对一些问题进行探讨并提出展望。     

In vitro embryonic axis and seedling shoot tip culture of Juglans nigra L.

Embryonic axes and seedling shoot tips of Juglans nitra L., Black walnut, were cultured in vitro. Significant variation existed among progeny from individual trees for growth of radicles and epicotyls and production of callus and axillary shoots from embryonic axes. The concentration of 6-benzyladenine influenced the growth of the radicle and epicotyl and production of callus and axillary shoots of axes. Axes generally initiated growth quicker on solidified woody plant medium than on Driver and Kuniyuki's walnut medium, but axillary shoot proliferation and elongation were eventually better on liquid Driver and Kuniyuki's walnut medium than on woody plant medium which required an etiolation treatment for microshoot elongation. The concentration of BA also influenced both callus growth and axillary shoot proliferation from seedling shoot tips. Axillary shoots which formed in Driver and Kuniyuki's walnut medium rooted best in sterile vermiculite following a 15 s dip in 10 mM indole-3-butyric acid. Micropropagated plants are growing in the greenhouse.     

云南野生稻不同染色体组型和外植体材料的离体培养研究

云南野生稻不同外植体愈伤组织诱导能力差别较大。花粉培养中愈伤组织诱导率差异在0％～11．8％之间,用成熟胚诱导愈伤组织,其诱导率在18．0％～35．2％之间,茎叶培养则在12．0％～25．0％之间。云南野生稻不同外植体诱导的愈伤组织再分化为绿苗的分化率在8．3％～100．0％之间。疣粒稻组培特性最好,东乡普通野生稻和景洪普通野生稻次之,药用稻最难组培。本文建立了疣粒、东乡、景洪普野3种野生稻的离体无性系,为长期保存云南野生稻资源奠定了基础。