拉曼光谱结合支持向量机对猪胞质内单精子显微注射胚胎性别的鉴定分析

【目的】 探求一种无损的、非侵入性的、快速的胚胎性别鉴定方法。【方法】 以207个猪卵胞质内单精子显微注射(intracytoplasmic sperm injection,ICSI)胚胎及其培养液为研究对象,单个胚胎经巢式PCR扩增鉴定其性别后随机分成训练集和测试集,同时利用拉曼光谱技术获取单个ICSI胚胎培养液的拉曼光谱。原始光谱经过处理后,采用支持向量机(support vector machine,SVM)算法构建分类模型,先用训练样本进行训练和建模,然后预测测试样本的性别,结合PCR性别鉴定的结果,计算分类准确率。【结果】 通过巢式PCR对207个胚胎进行性别鉴定,鉴定出71个雄性胚胎、128个雌性胚胎,8个样品扩增失败。猪ICSI雄性胚胎培养液在1 082和1 360 cm^-1拉曼位移处特征峰强度明显高于雌性胚胎,构建的胚胎性别鉴定模型的分类准确率为81.5%,雌性和雄性胚胎的分类准确率分别为81.3%和81.8%。【结论】 本研究将拉曼光谱技术与SVM法相结合构建了胚胎性别鉴定模型,分类准确率达到81.5%,提供了一种新的、无损的胚胎性别鉴定方法。     

湖羊胚胎和精液超低温保存效果的评价

【目的】 检验长期保存在国家家畜基因库的湖羊冷冻胚胎和冷冻精液的质量,评价超低温冷冻保存技术保种的效果。【方法】 对保存于国家家畜基因库的冷冻胚胎（保存20年）和冷冻精液（保存20和30年）进行复苏,进行相应鉴定后分别进行胚胎移植和人工授精,同时以0年冷冻精液和新鲜精液为对照,测定其受胎率和后代的羔皮性能、生长性能及繁殖性能,检验保存效果。【结果】 本试验共解冻胚胎36枚,其中,A级胚胎22枚,B级胚胎7枚,C级胚胎5枚,D级胚胎2枚,冷冻胚胎复苏利用率达到94.44%（34/36）,A级胚胎率达到61.11%（22/36）,移植34枚,产羔17只,冷冻胚胎复苏移植产羔率达50.00%;保存30年的冷冻精液复苏后平均精子活力达35%,受胎率为58.57%,平均产羔1.90只;保存20年冷冻精液复苏后平均精子活力达31%,受胎率为53.66%,平均产羔1.95只;胚胎复苏移植羊、30和20年冷冻精液后代体型外貌鉴定均符合纯种湖羊特征,没有畸形个体;羔羊一级羔皮率分别是31.25%、42.03%和23.81%,保持了当年湖羊的羔皮特性;生长发育及繁殖性能与现有湖羊群体性能基本保持一致。【结论】 利用超低温冷冻技术长期保存湖羊冷冻胚胎和冷冻精液的方法是可行的,对地方品种保种具有重要意义。     

马尾松胚胎发育的观察研究

在自然条件下，马尾松成熟花粉粒发育需6个月，授粉有效期在珠鳞展开的头5天，授粉后雌配子体游离核时期持续12个月，从游离核到细胞约需7天，颈卵器2－3个，受精作用发生在授粉后第13个月，原胚期约30天，幼胚期约40，成熟胚期约60天；从球花芽分化到种子成熟约需24个月，在形态发育的观察中，发现低温冻害会引起花粉在发育各阶段的形态异常及大量败育。     

离子注入对桉树萌发及苗期生长的影响

对6个尾叶桉和巨桉的全同胞和半同胞家系进行种子离子注入后,观察萌发和苗期生长,结果表明:(1)离子注入对种子发芽有刺激作用,既促进也抑制;(2)离子注入促进树高和侧根数生长,而抑制地径、叶长、叶宽和根生长,表明通过注入有可能获得生产中所需要的目的性状或品种;(3)离子注入的各家系平均生物量低于对照,但差异不显著;离子注入的家系平均叶比重、叶体积、叶肉体积和叶含水量均低于对照;(4)按突变临界值筛选,离子注入苗中共33株入选,突变率为5.323%;按各家系分别筛选,突变率在7.50%~30.00%.     

樟子松成熟胚的离体培养与不定芽的形成

沙藏45d的樟子松种子,剥胚培养在5个系列17种培养基上.在6BA0.2～2.0(mg/L,单位下同)+NAA0.05～0.5以及6BA1.0+IAA0.2～0.5的激素配比范围内,可诱导胚形成不定芽.KT与NAA或IAA配合时无不定芽形成.不定芽分化部位明显受激素种类控制,6BA与NAA配合可诱导胚轴分化出不定芽,而6BA与IAA配合,则可从子叶顶端和基部诱导出不定芽。     

Cross Breeding of Populus and Its Hybrids for Cold Resistance

Populus tomento a was crossed with P. tremuloidis, P grandiden ata, P. lba × P. grandidentata and P. alba × Ulmus s . ta pumila in order to maintain its rapid growth and high wood quality and improve its resistance to cold. Two methods were used to increase the germination rate from 1.5% to 41.1% and the remaining rate from 1.7% to 44.2%. Forty crossing combinations were conducted and 2 744 hybrid seedlings were obtained. MX. t4 × P grandiden ata (G-1-58), MX3 × P. tremuloidis (T-44-60), MX2 × P. tremuloidis (l-13-87-37) and MX2 × (P. alba × P. grandidentata) were regarded as superior combinations after analysis and selection. Thirty seedlings of these combinations and 11 triploid seedlings identified by counting their chromosomes were selected as super plants.     

桂花幼胚培养及愈伤组织增殖诱导

采用不同培养基对桂花胚的培养进行了试验研究,结果表明:MS 1-1.5 mg.L-16-BA有利于桂花幼胚的萌发;MS 2 mg.L-16-BA 0.3 mg.L-12,4-D有利于幼胚苗的幼叶愈伤组织的诱导,诱导率高,且质地致密;B5 1.5 mg.L-16-BA 0.05 mg.L-1NAA有利于幼胚苗基部产生的愈伤组织的增值诱导,所形成的新愈伤组织呈淡绿色,且质地致密,生长很好,并能分化出根。     

Tissue culture micropropagation of Douglas-fir

A procedure was developed for plantlet production from embryos of Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco]. The Medium for Conifer Morphogenesis, used at full strength, and supplemented with 10 M benzyladenine for 17 days produced an average of 6.8 shoots on more than 90% of the embryos. The percentage of shoot-forming embryos as well as the average number of shoots per embryo varied significantly among eight seedlots. For secondary multiplication, 89% of the adventitious shoots produced axillary buds on MCM with 5 M benzyladenine. However, 0.5 M BA was more suitable for the elongation of axillaries. Rooting ranged from 0–87% depending upon the treatment. The highest percentage was obtained with a 7-week incubation in peat:perlite containing 1/5-strength medium, 1% sucrose and 2.7 M naphthaleneacetic acid, followed by 5 weeks on peat:perlite with 1/5-strength major and minor salts, full iron and organics, 1% sucrose and 0.1% charcoal.