组织培养

Culture tender leaves in culture medium of MS+2,4-D1.5mg/L+30g/L suger+0.7% agar pH5.8 for 20 days in darkness at 25℃, and then subculture to induced Ⅱ-type calli. Use forceps cutting the tissue to nubble with 2mm2, and put the tissue into Agrodacterium tumefaciens LBA4404 liquid supplemented with AS 100mg/L,then, co-culture 3 days, resume 7 days in MS+2,4-D1.5mg/L+30g/L suger+500mg/L cef, take to MS+2,4-D1.5mg/L+30g/L suger+100mg/L cef+10.0mg/L kanamycin culture 20 days in darkness. After that to make it polarize in MS+30g/L suger+100mg/L cef+10.0mg/L km. The percentage of km resistant callus was reached max after infection for 45 min, the average is 29.66%. Simultaneity, tender leave callus are infected 5 min by A. tumefaciens liquid in different negative pressure, and kept on 15 min in Agrodacterium tumefaciens liquid without negative pressure. Then screen out resistant callus and obtain transgenic plants. When the negative pressure is -0.05MP the percentage of km resistant callus was reached max, the average rate is 37.5%.

将心叶接种于MS+2.4-D1.5mg/L+30g/L 蔗糖，琼脂0.7%，pH5.8 培养基中25℃暗培养20d 后继代一次，诱导产生Ⅱ型胚性愈伤组织，用镊子将其夹碎成2mm2大小的小块，置入添加100mg/L AS 的根癌农杆菌LBA4404 菌液中，侵染时间为45min，共培养3 天后，转入MS+2.4-D1.5mg/L+30g/L 蔗糖+500mg/L Cef 培养基中恢复7天，再转入MS+2.4-D1.5mg/L+30g/L 蔗糖+100mg/L Cef+10.0mg/L Km 中，遮光培养20d后，置入其MS+30g/L 蔗糖+100mg/L Cef+10.0mg/L Km 分化，卡那霉素抗性愈伤组织获得率在侵染45min 时达到最大值平均为29.66%；同时以甘蔗心叶愈伤组织材料，通过循环水式真空泵，产生负压，设定不同的负压值，在农杆菌菌液中感染5min 后，在负压条件下继续侵染15min 后，筛选出抗性愈伤组织并获得转化植株，其中在负压为-0.05 时转化率达到最大值，其卡那霉素抗性愈伤组织获得率平均为37.5%。

Establishment of a bovine mammary tissue culture system Bovine mammary tissue was cultured in superfusion system or stationary system, and the influence of these two methods in the activity and ultrastructure of tissue was investigated according to LDH vigor, trypan blue dying, agrose gel electrophoresis, transmission microscope observation.

1奶牛乳腺组织体外培养方法的建立采用组织灌流和组织块贴壁法体外培养奶牛乳腺组织，通过LDH活力测定、台盼兰染色、琼脂糖凝胶电泳、透射电镜技术比较两种培养方法对奶牛乳腺组织活性及组织超微结构的影响，建立奶牛乳腺组织的培养体系。

To establish a model of piglet testis tissue cultured in vitro,the testis's tissues of 30-day-old piglets were used to observe in vitro spermatogenesis,and the differentiation of androgone was identified pre- and post-culture by transmission electron microscopy and HE.

以30日龄仔猪睾丸组织块为培养对象，应用飞利普TECNAI10电子显微镜对培养前后的生精细胞类别进行了观察，对培养后的组织块进行生精细胞分化程度鉴定，建立了仔猪睾丸组织生精细胞爬片体外培养模型。

Plantlet regeneration from cotyledon of Citrullus lanatus cv. Zhangkang No. 4 was studied. The results showed that aseptic seedlings should be cultured in dark for 3d, then exposed to a photoperiod of 16/8h/d for 3d. The callus induction rate was higher in cotyledons placed facing downwards to the medium than in those facing upwards. The highest induction rate occurred in MS medium containing 6-BA (2.0mg/L), kinetin (1.0mg/L)and GA3 (1.0mg/L). The induction initiated in dark and took 7 days, while callus growth and differentiation proceeding for 7 days under 16/8h light-dark cycles. A highest rate of embryogenic callus was obtained after 3 successive subcultures.

以无子西瓜郑抗4号无菌苗子叶为外植体，进行了体细胞胚发生及植株再生的研究，结果表明，无菌苗应先进行3d暗培养，然后采取光照16h/d和黑暗8h/d培养3d；将子叶的叶面朝下放置于培养基上的愈伤组织诱导率高于叶面朝上的培养方式；子叶诱导胚性愈伤组织的最适培养基配方为MS+6-BA2.0mg/L+KT1.0mg/L+GA31.0mg/L；诱导需要在黑暗条件下启动，进行7d暗培养，而生长分化于光照16h/d和黑暗8h/d条件下培养7d；继代3次得到的胚性愈伤组织最多；最适生根培养基配方为1/2MS+IBA0.3mg/L。

In order to understand these differences on gene transcript level, even to get the genes and their expression related to cell culture, we researched the differences of total mRNA of these tissues by RAPD-PCR, the result showed that the difference between embryo and head of nymphae was much closer than embyo with others, which was accorded to the time order of insect upgrown, and the tissue which is more different from embryo on transcript level ismore diiBcult to be cultured in vitro.

四种组织的培养难易程度和分离出的细胞形态的不同，说明存在着组织表达的差异，为了从转录水平上来认识它们的差异，甚而获得与细胞培养难易相关的基因及其表达产物，本实验还采用了RAPD分析的方法进行了这四种组织：中后期虫卵胚胎、若虫头部组织、成虫的精巢和卵巢的mRNA表达差异研究，结果表明：从发育方向上看，四种组织的表达差异有分化越大差异系数越大的趋势，而在相同条件下则越难培养。

ABSTRACT It is because that the excellent acceptors for maize transgenic engineering are insufficient in our country, especially in the southwest mountain areas of china and hereditary variation regularity for the two characters such as efficiency of embryonic callus induction and number of regenerating plant (these two characters were abbreviated to the nduction efficiency and number of regenerating in the following of the paper, respectively), which hint the maize culturing capacity, is not very clear. Therefore, aiming at picking out superior acceptors, we had made systematic researches on the two characters with combing traditional quantitative-character genetic analyzing methods such as single-factor genetic mating design, diallel crossing genetic design, genetic effect analyzing method and the modern molecular locating method such as QTLs'. The main results are followed.(1) 50 superior inbred lines and about 30 crosses in our country, especially in the southwest of China were used for identifying and selecting the superior genotypes in the above two investigated characters under the same culturing condition in 2000 and 2001. There was very significant difference among the genotypes in the both characters. But the two characters were not certainly related. Some genotypes such as 18-599 and 18-599 were very good in them. For some ones such as zong 31, induction was higher than 18-599 and 18-599 in the efficiency, but it was only 1/3 to the later in regenerating number. In some genotypes such as S37, R08, R15, P138, A318, induction efficiency was just about 3% and scarcely any regenerating plants were got. On the whole, hybrids acted better than inbreeds in the both characters.(2) Two kind of inbreeds were selected as parents of the Griffing's method 1. 18-599 and 18-599 and the inbred line zong 31 are one kind because they are not only superior in the characters of maize cross breeding, such as CA, resistance to disease and the important agricultural characters, but also excellent in transformation characters as the induction and regeneration.

针对我国、特别是西南山地所需玉米转基因工程育种优良受体极为匮乏和反应玉米幼胚培养能力的2个主要性状，即玉米幼胚胚性愈伤组织诱导率和胚性愈伤组织绿苗发生数的遗传变异规律十分不清楚的实际情况，本研究从筛选玉米转基因工程所需要的优良受体入手，采用单因素遗传交配设计、双列杂交遗传交配设计、世代基因效应等传统数量性状分析方法，以及现代分子标记定位主效QTL分析方法，对玉米幼胚胚性愈伤组织诱导率和胚性愈伤组织绿苗发生数等2个性状进行了较为系统的分析研究，取得以下主要研究结果：(1)于2000年和2001年通过对我国、特别是西南地区近50份优良自交系和近30个杂交组合，在相同培养条件下，对幼胚培养胚性愈伤组织诱导率和胚性愈伤组织绿苗发生数等2个幼胚培养能力性状进行了筛选与鉴定，发现玉米不同基因型具有完全不同的幼胚培养胚性愈伤组织诱导率和胚性愈伤组织绿苗发生数，但幼胚培养胚性愈伤组织诱导率与胚性愈伤组织绿苗发生数并不具有必然的相关关系，有的基因型，如自交系18-599和18-599在胚性愈伤组织诱导率和愈伤组织绿苗发生数等2个性状都表现相当优异；有的基因型，如自交系综31，仅幼胚培养胚性愈伤组织诱导率性状表现高于19-599和18-599，但在胚性愈伤组织绿苗发生数这一性状则与它们有相当大的差距，仅为19-599和18-599的1/3左右；有的基因型，如S37、R08、R15、P138、A318等玉米自交系不仅幼胚培养胚性愈伤组织诱导率很低，平均仅在3％左右，而且胚性愈伤组织绿苗发生数表现也很差，基本上没有分化成苗。

This study was conducted to examiune the fibrotic effect of Ni-Ti and 317L al loys in esophagus.The extract fluid from Ni-Ti,317L alloys was made according t o the ASTM standards of U.S.A. The Fb of esophageal scar was cultured primarily ,then incubated with alloy abstract fluid. The proliferating activity of Fb was measured by MTT at 4, 24, 48, 72 hours in the course of culturing. The esophagu s embedding test of Ni-Ti,317L alloys was made according to ASTM standards of U .S.A.The tissue around the alloys was taken at weeks 2 and 12,and the pathologi c changes were analysed.

为探讨新型支架材料Ni-Ti、317L合金在食管局部的致纤维化作用，按美国ASTM标准制备NiTi、317L合金的金属浸提液；&组织块培养法&原代培养食管壁疤痕的成纤维细胞，传代后以金属浸提液进行培养，分组后分别培养4、24、48、72 h，MTT法检测不同培养时间后Fb增殖功能的变化；按美国ASTM标准进行NiTi、317L合金试件的食管壁内包埋实验，即将金属试件经表面处理后直接置入食管壁粘膜层与肌层之间，术后2、12周取出包埋组织，分析试件周围组织的病理变化，并进行胶原纤维染色，观察纤维形成状况。

This page provides some protocols about porcine brain tubulin, sea urchin sperm axonemes, rat liver cell cytosol and organelle fractions, organelle motility assay, tissue culture of PtK1 cells, basic protein chemistry and so on.

该网页提供了一些有关猪脑的微管蛋白、海胆的精子基因丝、大鼠肝脏细胞的细胞质和细胞器的级分、细胞器的运动性实验、PtK1细胞的组织培养、基本的蛋白化学等等方面的操作流程。

American black walnut ; tissue culture ; Xinjiang Autonomous Region

美国黑核桃；组织培养；新疆

There is a badly explant browning during the process of potato tissue culture.

［目的］在马铃薯的组织培养过程中存在着严重的外植体褐化现象。

It has been put forward that there exists single Ball point and double Ball points on the symmetrical connecting-rod curves of equilateral mechanisms.

从鲍尔点的形成原理出发，分析对称连杆曲线上鲍尔点的产生条件，提出等边机构的对称连杆曲线上有单鲍尔点和双鲍尔点。

The factory affiliated to the Group primarily manufactures multiple-purpose pincers, baking kits, knives, scissors, kitchenware, gardening tools and beauty care kits as well as other hardware tools, the annual production value of which reaches US$ 30 million dollars.

集团所属工厂主要生产多用钳、烤具、刀具、剪刀、厨具、花园工具、美容套等五金产品，年生产总值3000万美元，产品价廉物美、选料上乘、质量保证，深受国内外客户的青睐