短叶对齿藓组织培养的研究

以采自白云鄂博主矿区的短叶对齿藓为试验材料,研究了不同消毒剂以及不同浓度植物激素6-BA和IAA对短叶对齿藓愈伤组织诱导和分化的影响。结果表明:短叶对齿藓配子体最佳消毒剂及作用时间为体积浓度百分比75%乙醇浸泡30s,再用质量浓度0.1g/L升汞消毒90s;采用Knop培养基培养短叶对齿藓茎叶段,质量浓度为0.1mg/L 6-BA促进愈伤组织分化形成配子体,质量浓度1.0mg/L 6-BA则抑制短叶对齿藓愈伤组织形成,IAA有助于原丝体的萌发生长。     

尖叶拟船叶藓原丝体发育特征研究

将尖叶拟船叶藓[Dolichomitriopsis diversiformis(Mitt.)Nog.]孢子接种于Knop培养基上,置于恒温培养箱中培养,在光学显微镜下对其原丝体(protonema)发育特征进行了详细观察和记录。结果表明:孢子第2天就开始萌发,第6天时其萌发率达90%以上;原丝体系统由绿丝体(chloronema)和轴丝体(caulonema)构成,假根(rhizoides)产生于芽体基部,由轴丝体退化而成;配子枝原始细胞产生于绿丝体分枝的基部或轴丝体上的斜壁细胞;配子枝(game tophore)形成后其上各部位都可形成假根;孢子萌发类型为真藓型(Bryum-type)。     

红蒴立碗藓愈伤组织的诱导和植株再生

1植物名称红蒴立碗藓（Plryscomitriun eurystomum Sendtn.）,也称尖口立碗藓、广口立碗藓。2材料类别配子体。3培养条件脱毒培养基：Knop（张献龙和唐克轩2004）;愈伤组织诱导培养：MS＋6-BA0．05mg·L-1;再分化培养基：MS。以上培养基均添加2％葡萄糖和1％琼脂。培养温度为（25±1）℃,光照强度为60-80μmol．m-2．s-1,光照时间为12h．d-1。4生长与分化情况4．1无菌材料的获得对采集得到的配子体植株先用75％酒精溶液消毒5S,无菌水清洗1次;再用0．25％新沽尔灭溶液表面消毒10S,无菌水漂洗3次;在无菌滤纸上将材料水分吸干,接种到不添加任何激素的Knop培养基上,培育获得无菌材料（图1）。     

仙鹤藓的孢子萌发及愈伤组织诱导

通过对采自河北雾灵山海拔1500m的仙鹤藓（Atrichum undulatum）的孢子萌发以及原丝体发育的观察,发现仙鹤藓孢子无休眠现象,孢子接种3天左右萌发：其原丝体发育分为绿丝体和轴丝体两个阶段。扩大培养实验结果表明。仙鹤藓茎叶体在添加2％葡萄糖的MS培养基上,置于25℃／20℃、14小时光照/10小时黑暗、36μmol·m^-2·s^-1条件下培养．产生新生茎叶体最多,且茎叶体长势最好,可以获得大量无菌材料。仙鹤藓愈伤组织诱导实验显示,形成愈伤组织的最佳培养基为添加2％葡萄糖和1．0mg·L^-16-BA的MS培养基。     

不同培养基及酶对立碗藓原丝体的作用研究

采用含不同成分的MS培养基处理立碗藓的原丝体,发现随蔗糖浓度的提高原丝体的生长相对趋于老化;当培养基中附加不同浓度的2,4-D和6-BA 时,10日龄的原丝体出现了明显变化,当二者浓度均为0.5 mg*L-1,幼嫩的原丝体上能长出疏松易碎的白绿色愈伤组织.利用1%纤维素酶和0.8%的果胶酶分别处理10日龄、20日龄的原丝体和10日龄的茎叶体的叶片2～10 h,结果发现10日龄的原丝体在酶解2 h后能游离出大量的原生质体;而20日龄的原丝体效果较差,只能游离出极少量的原生质体;对于10日龄的茎叶体酶解10 h仍看不到原生质体游离出来.这表明立碗藓原丝体对不同的外界条件可产生不同的反应,但原丝体的生理状态不同对激素和酶类的反应具有非常显著的差别.     

光照和温度对尖叶拟船叶藓孢子萌发及原丝体发育的影响

用组织培养法和光学显微镜技术初步研究了光照和温度对尖叶拟船叶藓孢子萌发及原丝体发育的影响。结果表明：（1）光照是影响孢子萌发的主要环境因子,20℃环境下,24h光照4d的孢子萌发率达83.3％;温度下黑暗培养的孢子30d也不能萌发,转光照后4d的萌发率可达84.6％;（2）温度是影响原丝体发育的主要环境因子,连续光照下,20℃的原丝体生长最快、分枝最多、分化最早,第31天可长达651.64μm;25℃次之,只有379.12μm;而自然光照下5-10℃环境下的孢子萌发率（18d为70.2％）和原丝体生长速度（127.44μm）均最慢;（3）原丝体发育到一定阶段,断裂为单个细胞,单个细胞再萌芽出新原丝体。     

东亚砂藓组织培养技术方法研究

以东亚砂藓（Racomitrium japonicum）配子体为外植体,比较了不同消毒液对消毒效果的影响,对接种消毒外植体的培养基进行了尝试性筛选,并研究了蔗糖浓度对东亚砂藓配子体生长的影响。结果显示,将东亚砂藓配子体用2％洗洁精溶液浸泡数分钟后,再用0．02％的升汞处理45-60S的消毒效果最佳;有机培养基是无菌外植体接种的最佳培养基;3％的蔗糖更有利于无菌外植体产生的原丝体团和幼嫩配子体的生长。     

腾格里沙漠固定沙丘藓类植物结皮层的自然恢复及人工培养试验研究

 通过藓类结皮层的自然恢复和人工培养藓类植物促进结皮层形成试验,研究了腾格里沙漠固定沙丘生物结皮层形成过程中优势成分真藓 （Bryum argenteum） 的繁殖生物学特性,结果表明：藓类结皮层人工去除后在3～4年内70%的样方基本恢复,在此过程中真藓主要靠茎叶碎片传播和繁殖;通过分株法、撒茎叶法培养的真藓在1个月后长满整个样地, 主要通过如下方式繁殖——茎的碎片连续分枝可产生小植株,茎、叶均可产生原丝体,由原丝体发育产生小植株,小植株又可再生原丝体,如此反复产生新植物体,这一过程是野外人工促进生物结皮层形成过程中真藓主要的繁殖途径。与室内培养中真藓的繁殖特性相比较,野外培养的真藓在繁殖过程中产生的原丝体较粗壮,分枝多,但在两种条件下的繁殖特性相同,能够揭示该地区自然条件下藓类植物萌发和定居的繁殖机理。该研究为人工促进生物结皮层形成及治理受损结皮层提供了实验依据。     

孢子大小和培养基对水蕨孢子萌发及配子体性别分化的影响

利用MS培养基、改良Knop’s培养基、自来水和蒸馏水分别培养水蕨中等大小孢子,同时利用改良Knop’s培养基培养不同大小的水蕨孢子,观察记录不同条件下水蕨孢子萌发和性别分化情况。实验表明,二级孢子（赤道轴120～140μm）萌发率最高;一级孢子（赤道轴〉140μm）萌发最有利于使水蕨发育为两性配子体,三级孢子（赤道轴〈120μm）萌发最有利于使水蕨发育为雄配子体;MS培养基和改良Knop’s培养基相对于自来水和蒸馏水有利于水蕨孢子萌发;各培养基中水蕨两性配子体比率排序是MS培养基〉改良Knop’s培养基〉自来水〉蒸馏水,而雄配子体比率排序与之相反。此结果为水蕨的引种保护、人工繁育和分子生物学研究提供理论依据。     

直齿藓属一新组一新种

报道真藓科直齿藓属一新种,具边直齿藓Ｏ．ｂｉｌｉｍｂａｔｕｍＸ．Ｊ．Ｌｉ ｅｔＤ．Ｃ．Ｚｈａｎｇ。本种主要特征（１）蒴齿双层,内齿长,狭线形,无基膜。（２）叶边缘明显分化,具２￣３列狭长细胞,除近尖部和基部边缘外,明显两层。（３）叶细胞明显宽于本属各种（宽达１８￣２４μｍ）。（４）蒴盖短圆锥形,无长喙状尖;本种介于Ｏｒｔｈｏｄｏｎｔｉｕｍ与Ｏｒｔｈｏｄｏｎｔｏｐｓｉｓ两属之间,根据蒴齿双层之重要特     

水青树组织培养中无菌苗培养条件的优化

对水青树组织培养中种子的消毒方法和无菌苗培养条件进行了优化。4种消毒方法的效果比较表明,0.1% HgCl₂溶液对水青树种子的发芽有较强的抑制作用,而4% NaClO溶液的消毒效果最佳,对发芽的抑制作用最小。不同培养基上水青树种子的发芽情况显示,中盐低氮培养基有利于种子萌发,完全1/4 MS培养基为水青树无菌苗培养的最佳培养基。     

Effects of chemical substances on the rapid cultivation of moss crusts in a phytotron from the Loess Plateau,China

In this study, typical moss crusts, which were dominated by the species Didymodon vinealis (Brid.) Zand., were collected from the Loess Plateau and a 65-day cultivation experiment was performed to study the effects of five kinds of nutrient solutions (Knop, Murashige-Skoog (MS), Benecke, Part and Hoagland), two kinds of carbohydrates (glucose and sucrose) and three kinds of plant growth regulators (thidiazuron (TDZ), 6-benzylaminopurine (6-BA) and naphthaleneacetic acid (NAA)) on the coverage, plant density, and plant height of moss crusts. The main conclusions are as follows. (1) All Knop, MS, Benecke, Part and Hoagland nutrient solutions improved the coverage and plant density of moss crusts to different degrees and the promotional effects of the Hoagland nutrient solution were most significant. (2) Glucose and sucrose could promote the formation of moss crusts, but they inhibited the development of moss crusts at concentrations greater than 10?g/L. (3) With an increase in concentration, the effects of TDZ on the development of moss crusts changed from “enhanced” to “inhibited”. Regardless of whether the concentration was high or low, 6BA had no significant effects on the growth of moss crusts, and NAA reduced the development of moss crusts. Results suggest that nutrient solutions (e.g. Hoagland), low concentration carbohydrates solutions, and some plant growth regulators (e.g. 1?mg/L TDZ) enhance the development of moss crusts in Loess Plateau under the appropriate environmental conditions.     

粗茎鳞毛蕨孢子萌发研究

以经过3年低温储藏的粗茎鳞毛蕨孢子为实验材料,从孢子离心、孢子消毒、培养基种类、光质等4方面对孢子萌发进行研究,结果表明:在离心转数≤14 000 r.min-1、离心时间≤30 min条件下,离心处理对孢子萌发基本无影响;对孢子进行1%NaClO水溶液浸泡处理20～30 min为最佳消毒条件;改良Knop’s培养基为最佳孢子萌发培养基;黑暗条件下孢子不能萌发,但是黑暗处理能够明显提高孢子萌发整齐性;红光比白光能促进孢子提早萌发1 d左右,但对提高萌发率效果不显著。     

Large-scale analysis of 73 329 physcomitrella plants transformed with different gene disruption libraries: production parameters and mutant phenotypes

Gene targeting in the moss Physcomitrella patens has created a new platform for plant functional genomics. We produced a mutant collection of 73 329 Physcomitrella plants and evaluated the phenotype of each transformant in comparison to wild type Physcomitrella. Production parameters and morphological changes in 16 categories, such as plant structure, colour, coverage with gametophores, cell shape, etc., were listed and all data were compiled in a database (mossDB). Our mutant collection consists of at least 1804 auxotrophic mutants which showed growth defects on minimal Knop medium but were rescued on supplemented medium. 8129 haploid and 11 068 polyploid transformants had morphological alterations. 9 % of the haploid transformants had deviations in the leaf shape, 7 % developed less gametophores or had a different leaf cell shape. Other morphological deviations in plant structure, colour, and uniformity of leaves on a moss colony were less frequently observed. Preculture conditions of the plant material and the cDNA library (representing genes from either protonema, gametophore or sporophyte tissue) used to transform Physcomitrella had an effect on the number of transformants per transformation. We found correlations between ploidy level and plant morphology and growth rate on Knop medium. In haploid transformants correlations between the percentage of plants with specific phenotypes and the cDNA library used for transformation were detected. The number of different cDNAs present during transformation had no effect on the number of transformants per transformation, but it had an effect on the overall percentage of plants with phenotypic deviations. We conclude that by linking incoming molecular, proteome, and metabolome data of the transformants in the future, the database mossDB will be a valuable biological resource for systems biology.     

Tight control of growth and cell differentiation in photoautotrophically growing moss (<Emphasis Type="Italic">Physcomitrella</Emphasis><Emphasis Type="Italic">patens</Emphasis>) bioreactor cultures

The use of the moss Physcomitrella patens as a production system for heterologous proteins requires highly standardised culture conditions. For this purpose a semi-continuous photoautotrophic bioreactor culture of Physcomitrella was established. This culture grew stably for 7 weeks in a 5-l bioreactor with a dilution rate of 0.22/day. Enrichment of the air for aeration in a batch bioreactor culture with 2% (v/v) CO₂ resulted in an increase in the specific growth rate to 0.57/day. Changes in the pH of the semi-continuous bioreactor culture medium between pH 4.5 and pH 7.0 influenced protonema differentiation; however it did not negatively affect the growth rate compared to uncontrolled pH. The advantages of Physcomitrella as a system for the production of heterologous proteins in plants are discussed.     

梨蒴珠藓愈伤组织诱导和配子体再生研究

以梨蒴珠藓无菌藓株为外植体诱导愈伤组织和配子体再生,接种于含不同激素组合的MS和Knop固体培养基上,分别进行愈伤组织和不定芽的分化,并探讨愈伤组织诱导和配子体再生的适宜培养条件.结果显示,愈伤组织诱导的最佳培养基是MS+0.5 mg/L BA+0.1 mg/L 2,4-D,愈伤组织诱导率为33.3%;不定芽诱导的最佳...     

橙黄玉凤花种子萌发培养及小苗组培快繁研究

对橙黄玉凤花Habenaria rhodocheila种子进行无菌萌发培养以获得大量原球茎,并对原球茎组培,建立其快繁体系。结果表明,橙黄玉凤花种子以0.1%升汞消毒15 min为最佳处理。种子萌发培养基中,添加0.2%活性碳(AC)有利于种子萌发。种子萌发的最佳培养基为1/2MS＋NAA 0.5 mg·L-1＋AC 0.2%,原球茎增殖分化最佳培养基为1/2MS＋NAA 0.2 mg·L-1＋ZT 3.0 mg·L-1＋AC 0.2%;生根最佳培养基为1/2MS＋NAA 0.1 mg·L-1＋6-BA 2.0 mg·L-1＋AC 0.2%。     

垂丝海棠组培再生体系建立的研究

目的:通过对垂丝海棠(Malus halliana koehne)进行组织培养研究为木本观赏海棠提供技术支持。方法:采用常规组培手段及正交设计试验对外植体消毒时间、快繁培养、叶片愈伤诱导、再生、生根等进行研究。结果:外植体最佳消毒时间:75%酒精30s 0.1%升汞10min;最适增殖培养基为:MS 6-BA0.7mg/L NAA0.3mg/L,增殖系数可达7;正交设计结果显示激素对叶片愈伤组织诱导影响的因素大小为:2,4-D>6-BA>NAA;再生最佳配方为6-BA6.0mg/L NAA0.5mg/L再生效率达91.8%;最适生根培养基为:1/2MS IBA0.3mg/L 蔗糖20g/L。结论:对垂丝海棠组培再生研究取得初步成功。     

金灰藓(Pylaisiella polyantha)配子体发生的实验观察

金灰藓Pylaisiella polyantha (Hedw．)Grout孢子在光照培养箱中进行培养,实验组为含有Knop培养液的琼脂培养基质,对照组为无营养成分的琼脂培养基质。在光学显微镜下对配子体发生进行了观察、描绘和照相。结果表明：实验组和对照组孢子萌发所需时间相同,均为53．5h,萌发极相为1～4极。4d时,实验组萌发率为85．4％,对照组为54．1％;20d时,开始形成配子体芽原基;40d时,形成具假根、茎、叶的配子体,并形成两性器官。并对配子体发生过程的特点进行了分析和讨论。