激素对洋桔梗植株再生的影响及生根培养的研究

以MS为基本培养基 ,附加不同浓度的 6 BA、KT、NAA和IBA诱导洋桔梗叶片外植体的再生植株。结果表明 :MS +6 BA 0 .5～ 1 .0mg/L(单位下同 ) +NAA 0 .2和MS +6 BA 0 .5～ 1 .0 +IBA 0 .2培养基都能诱导外植体产生愈伤组织 ,但 6 BA的浓度必须小于 1 .0mg/L ,否则会导致组织的严重玻璃化 ;MS +KT 1 .0～2 .0 +NAA 0 .2或MS +KT 1 .0～ 2 .0 +IBA 0 .2培养基也能诱导外植体产生愈伤组织 ,愈伤组织出现的时间较早且质地较好 ,适合分化。继代培养时 ,MS培养基中仅加 6 BA 0 .5mg/L或KT 0 .5mg/L ,即能获得较高的分化率。生根培养研究中 ,培养液为 1 /2MS+5 0g/L糖 +IBA 2mg/L的前处理 ,生根效果较好 ,生根率接近基质生根培养的生根率。     

‘红阳’猕猴桃叶盘高频直接再生体系的建立

以‘红阳’猕猴桃雌株幼叶为外植体,直接诱导产生不定芽,并对不定芽增殖以及生根体系进行优化,建立了高频再生体系。结果表明,在MS+3.0 mg/L BA+1.0 mg/L NAA培养基中,不定芽诱导率达100％,平均出芽数达18.67个/叶盘;在MS+2.0 mg/L BA+1.0 mg/L NAA+0.1 mg/L GA3培养基中,增殖率达100％,1~6代不定芽平均繁殖系数达8.63;不定芽在1/2 MS+0.8 mg/L IBA培养基中培养15 d,然后继代至含1/2 MS液体培养基的珍珠岩中培养15 d,生根率达100％,且其根系发育良好;98株试管苗移栽到土壤盆钵中,成活95株,成活率达96.94％。本试验成功建立了红阳猕猴桃叶片高频直接再生体系,该体系诱导产生不定芽周期短,出芽率高,不定芽增殖系数大,生根率高且根系发达,为红阳猕猴桃试管苗的工厂化生产和遗传转化奠定了基础。     

野葛高频植株再生体系相关因子的优化

对野葛高频植株再生体系相关因子进行优化,结果表明,野葛带芽茎段再生体系的最佳消毒方式为70%酒精处理30s后用0.1% HgCl₂处理15min;最佳培养基为MS+NAA 0.5mg/L+6-BA 5.0mg/L;最佳培养条件为:蔗糖浓度30g/L,pH 6.0,液体培养,培养温度25℃,光照培养。     

中国传统菊花品种‘小林静’再生及转化体系的建立

以中国传统菊花品种‘小林静’叶片为外植体,建立了‘小林静’较好的再生体系及遗传转化体系.结果表明,‘小林静’叶盘最适不定芽分化培养基为MS+2.0 mg/L 6-BA+1.5 mg/L NAA,不定芽分化率为92.76％,平均再生不定芽数为2.3767个;试管苗最佳生根培养基为1/2MS,生根率达100％.移栽采用灭菌的蛭石,再生苗移栽成活率达90％以上.采用根癌农杆菌C58C1介导的叶盘转化法进行‘小林静’的遗传转化试验,农杆菌OD600=0.5-0.6,侵染10 min后,将叶片外植体接种到MS+2.0 mg/L 6-BA+1.5mg/L NAA的培养基中黑暗共培养2d,之后转接到附加10 mg/L硫酸卡那霉素和400 mg/L羧苄青霉素的分化筛选培养基中进行转化细胞的筛选,待长出抗性芽后转接至生根培养基中进行培养,最终建立了菊花品种‘小林静’的遗传转化体系.     

荞麦组织培养及高频植株再生体系的建立

通过对荞麦(Fagopyrum esculentum Moench)不同外植体、不同激素配比的比较研究,建立了荞麦离体培养高效植株再生体系。荞麦子叶切段在含2．0 mg/L 2,4-D和1．0 mg/L 6-BA的MS培养基上愈伤组织诱导率为89．6％,而下胚轴切段在含2．0 mg/L 2,4-D和1．0-2．0 mg/L 6-BA MS培养基上愈伤组织诱导率高达100％。在2．0 mg／L 6-BA、0．1 mg,L IAA和1 mg／L KT的MS培养基上通过愈伤组织间接分化或外植体直接分化形成不定芽。来自子叶和下胚轴的愈伤组织的分化率分别为42．5％和73．6％,下胚轴的分化率明显高于子叶。将生长状态良好的不定芽转至含1．0 mg/L IBA和0．5mg/L NAA的1/2 MS培养基上生根,生根率达到100％。再生植株移栽到盆土中,成活率达91．6％,并且生长状态和特征均表现正常。     

绵枣儿叶片植株再生体系的建立

1 植物名称 绵枣儿[Scilla scilloides（Lindl.） Druce]。     

荞麦高频离体再生及发根农杆菌转化体系的建立

荞麦无菌苗下胚轴切段在不同激素配比的MS培养基上诱导愈伤组织,出愈率均为100％。在2．0mg/L2,4-D和1．5mg/L 6-BA组合下诱导产生的愈伤组织;转入2．0mg/L 6-BA和1．0mg/L KT的MS培养基,再生苗分化率在80％以上。根尖色体分析表明再生植株具一定的遗传稳定性。发根农杆菌A4转化荞麦下胚轴和子叶获得发状根,纸电泳检测所有随机取样测定的发状根均有相应冠瘿碱的存在。     

‘贵长’猕猴桃叶片高效直接再生体系的建立

以‘贵长’猕猴桃叶片为外植体,直接脱分化产生不定芽,并对不定芽增殖以及生根体系进行优化,建立了其高效直接再生体系。结果表明,叶片在MS+4. 0mg/L 6-BA+0. 4mg/L NAA培养基中,不定芽诱导率达95. 8%,平均出芽数达15. 7个/叶片;不定芽在MS+3. 0mg/L 6-BA+0. 3mg/L NAA+0. 2mg/L GA3培养基中,增殖率达100%,且1～6代平均繁殖系数达8. 15;不定芽先在添加1. 0mg/L IBA的1/2 MS固体培养基中诱导7d,然后再先后在1/2 MS固体培养基和充分吸附1/2 MS培养液的珍珠岩中各培养14d,生根率达98. 61%,且根系发育良好; 50株试管苗移栽到以珍珠岩和田间土壤(其体积比为1∶4)为基质的营养钵中,2周后成活49株,成活率达98%。该研究成功建立了‘贵长’猕猴桃叶片高效再生体系,该方法不定芽诱导周期短,出芽率高且数目多,不定芽增殖系数大,生根率高且试管苗根系发达,为‘贵长’猕猴桃离体快速繁殖和遗传转化奠定了基础。     

‘巴斗’杏再生体系的建立与耐盐突变体的筛选

以‘巴斗’杏试管苗茎段为外植体,研究其再生体系的建立以及在含不同浓度NaCl培养基上诱导耐盐愈伤组织,筛选耐盐突变体。结果显示：茎段在MS＋6-BA1．5mg·L^-1＋IBA0．5mg·L^-1培养基上诱导愈伤组织效果最好,芽的分化率可达88％;将出芽愈伤组织块接种到附加IBA0．5mg·L^-1＋KT2．0mg·L^-1的MS分化培养基上效果最佳,芽的分化系数最高为12．7;较理想的生根培养基为MS＋NAA0．1mg·L^-1。＋IBA0．2mg·L^-1,生根率在46．3％以上;在含0．8％NaCl的愈伤组织诱导培养基中,连续继代筛选2代,转入不含NaCl的分化培养基中,分化出了完整植株。经继代培养筛选,测定发现获得的耐盐植株比正常培养植株的游离脯氨酸含量高。     

川芎高频再生体系的建立

为建立川芎(Ligusticum chuanxiong Hort.)高频再生体系,优化了诱导和分化培养基及培养条件.以叶柄为外植体,以MS为基本培养基,KT2.0 mg/L+ IAA0.5mg/L的激素组合对不定芽分化最有利.在此基础上,针对外植体来源、培养条件和愈伤组织继代时间3个因素进行优化.结果表明:采用川芎无菌苗叶柄作为外植体,黑暗条件下诱导出愈伤组织,再在光照下继代培养15d后转入分化培养基中对不定芽诱导最为有利,分化率为44.4％.分化后得到的不定芽在含NAA0.5 mg/L和IBA 0.5 mg/L的1/2MS培养基上生根率达90％,移栽存活率为95％.     

新疆杨高效遗传转化系统的建立

选择新疆杨(Populus alba L.var．pyramidalis Bge．)为遗传转化受体材料,为建立根癌农杆菌介导新疆杨高效遗传转化系统,从预培养时间、侵染时间、共培养时间、添加乙酰丁香酮(AS)的时机、共培养培养基中添加乙酰丁香酮浓度、侵染菌液的制备方法、外植体继代方式等7个方面优化筛选。结果显示较合适的转化系统为：预培养8h,农杆菌菌液(OD600=0．4)侵染15min,共培养5d,侵染菌液的最优制备方法是液体培养活化农杆菌2次加离心收集菌体重悬,共培养培养基中添加乙酰丁香酮80μmol／L。新疆杨叶盘转化频率可达38．10％。     

多年生黑麦草再生体系的建立

以多年生黑麦草‘高帽2号’(Lolium perenne‘Top Hat 2’)颖果为外植体,分别研究了胚端半粒颖果、纵切的胚端半粒颖果、胚端颖果小块、颖果小块这四种外植体处理方式与氯化汞、次氯酸钠两种表面灭菌方法对其愈伤组诱导的影响;利用两因素(2,4-D和6-BA)随机区组试验设计研究了不同植物生长调节物质及其浓度配比对外植体愈伤组织诱导的影响;利用三因素(NAA,6-BA和ZT)四水平正交试验设计研究了不同植物生长调节物质及其浓度配比对不定芽诱导的影响;研究了0.10 mg·L-1的NAA和IBA分别对不定根诱导的影响。结果表明:相同条件下,以饱满胚端半粒颖果为外植体,75%酒精表面灭菌1 min后,再用含1 m L·L-1Tween 20的次氯酸钠溶液(有效氯含量10%)处理30 min,愈伤组织诱导率最高;适宜的愈伤组织诱导培养基为MS+5 mg·L-12,4-D+0.03 mg·L-16-BA(p H=5.8),诱导率达68.7%,获得的高质量愈伤组织;不定芽诱导培养基为MS+0.5 mg·L-1NAA、0.5 mg·L-16-BA+0.9 mg·L-1ZT,不定芽诱导率最高,为56%;不定根诱导培养基为1/2MS+IBA培养基时,根系粗壮,诱导生根率为90%。该研究建立了多年生黑麦草高效组培再生体系,为高效基因工程育种的开展奠定了良好基础。     

草木樨状黄芪高频离体再生体系的建立

以草木樨状黄芪无菌苗茎切段为材料,在含１－２mg/L2,4-D和０－０．５mg/L６ＢＡ的MS培养基上培养获得大量愈伤组织,愈伤组织诱导率在９５％以上,愈伤组织在附加０．２mg/LＫＴ,１mg/L６ＢＡ,０或０．５mg/LＮＡＡ,５００mg/LＣＨ 和２００mg/L ＹＥ的ＭＳ培养基上诱导丛生芽,并进而发育成苗。苗的分化频率为１００％。分化苗或其茎的切段在不国源植物激素的１／２ＭＳ培养基上可出现根的分化,分化频率达９０％以上,再生植株经炼苗后移栽成活率达８０％以上。     

The determination of sensitivity parameters for auxin-induced H+-efflux from Avena coleoptile segments

Abstract A method is described for the measurement of auxin-induced H⁺-efflux from small populations of Avena coleoptile segments. The method allows the simultaneous investigation of the kinetics of rapid auxin responses over a wide concentration range. IAA promoted linear rates of H⁺-efflux and the change in amplitude of response occurred mainly over a low, narrow concentration range (10–50 μmol m^-3). The sigmoidal curve of best-fit to each set of dose-response data was determined using non-linear regression techniques, allowing the objective determination of characteristic tissue sensitivity parameters (R_MIN, R_MAX, K_D and p). The sensitivity parameters for the auxin-type herbicide fluroxypyr are also presented as well as IAA parameters determined in the presence of abscisic acid and the ‘antiauxin’ PCIB. The interpretation of the parameter values and the potential use of sensitivity parameter analysis for the evaluation of theories concerning plant hormone action and interaction is discussed.     

宁夏不同地区灵武长枣果实多糖的单糖成分分析

以宁夏4个不同地区(灵武、中宁、青铜峡、银川)成熟期的灵武长枣果实为研究对象,经水提醇沉法提取,采用DEAE-cellulose52和HW-55S分离纯化,并利用GC-MS法进行多糖的单糖组成分析。结果表明:多糖提取率最高的是灵武地区,达到1.795%;分离纯化后,4个地区的长枣多糖各得到1个中性(Ju-0)和3个酸性组分(Ju-1、Ju-2、Ju-3),其中Ju-2含量最高;GC-MS分析可知灵武长枣多糖含有阿拉伯糖、鼠李糖、核糖、岩藻糖、木糖、甘露糖、半乳糖、葡萄糖、葡萄糖醛酸、半乳糖醛酸10种单糖,不含果糖,以阿拉伯糖、核糖、半乳糖和2种糖醛酸为主,木糖含量最低。各地区多糖的单糖组成、含量各不相同,从各组分来看,四个地区多糖的Ju-0和Ju-1组分组成均以阿拉伯糖、核糖、半乳糖为主,四个地区多糖的组成差异主要在于Ju-2和Ju-3组分。从各地区单糖总量来看,灵武地区是阿拉伯糖含量最高,中宁、青铜峡、银川地区以葡萄糖醛酸含量为最高。     

Comparison of developmental stages of inflorescence for high frequency plant regeneration in Triticum aestivum L. and T. durum Desf.

Summary Whole immature inflorescences at 4 different developmental stages (0.5, 1.0, 1.5, 2.0 cm in size) of different genotypes of Triticum aestivum and T. durum were cultured to see the morphogenetic responses on Murashige and Skoog's (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) (2.5 mg/l). Very young inflorescences 0.5 and 1.0cm long formed embryogenic callus from their entire surface while 1.5 and 2.0 cm long inflorescences formed embryogenic callus from the basal spikelets and rachis only. This embryogenic callus was maintained by regular subcultures on MS medium with 2,4-D (2.5 mg/l) for more than a year. Plantlets were regenerated by transferring the embryogenic callus on hormone-free MS medium. Inflorescences (0.5 and 1.0 cm long) responded best in forming callus as well as plantlets at a very high frequency. Variation in response was observed amongst the genotypes but the qualitative response of formation of embryogenic callus and later regeneration of plantlets was observed from all the genotypes. Immature young inflorescence explants could provide a suitable material for particle gun mediated genetic transformation in wheat.Abbreviations BAP 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - MS Murashige and Skoog (1962)     

皖油离体培养再生植株体系的建立

甘蓝型油菜(Brassica napus L.)皖油14具柄子叶为外植体,进行安徽省地方性油菜品系离体培养再生植株体系的研究.通过不同浓度的激素组合实验,结果表明:在愈伤组织诱导中,以MS-C7(MS 2 mg/L6-BA 0.2mg/L NAA 0.25 mg/2,4-D 7.5 mg/L AgNO3)培养基的出愈率最高,且可产生大量绿色芽点;以MS-B3(MS 1.0 mg/L6-BA 0.1 mg/L NAA 7.5 mg/L AgNO3)培养基的芽苗分化率最高,可分化出丛生的不定芽;生根培养基MS 0.1 mg/L NAA 7.5 mg/L AgNO3的生根率达100%.7.5 mg/L的AgNO3可以明显减少愈伤组织的褐化,良好的空气流通可以减少玻璃苗的形成.初步建立了皖油14品系的高频再生植株体系.     

Rapid and high frequency shoot regeneration from hypocotyl protoplasts of Brassica nigra

Protoplasts, isolated from etiolated hypocotyls of seven day old seedlings of Brassica nigra, were cultured in Kao's liquid medium containing 7.2% glucose, 2,4-d (1 mg 1^-1), NAA (0.1 mg 1^-1) and zeatin riboside (0.5 mg 1^-1). After initial incubation for 3 days in dark at 25±1°C, cultures were transferred to a photoperiod cycle of 16/8 h and diluted on seventh and tenth day with MS medium containing 3.4% sucrose, 2,4-d (0.1 mg 1^-1) and BAP (1 mg 1^-1). About 62% of the cells divided at least once and 46% of them reached 8–16 cell stage in one week. The dividing cell clusters could be plated on agarose medium on the fifteenth day to obtain proliferating minicalli with a plating efficiency of 1.8%. 56.8% of minicalli, regenerated shoots on a regeneration medium containing 2 IP and IAA at 1 and 0.2 mg 1^-1 respectively. The in vitro produced shoots were rooted in MS medium containing 1 mg 1^-1 IBA and established in soil without difficulty. The time taken for protoplasts to develop into plants varied from 9 to 10 weeks.Abbreviations 2,4-d 2,4-dichlorophenoxyacetic acid - BAP 6-benzylaminopurine - 2 IP 2-isopentenyladenine - IAA indole-3-acetic acid - IBA indole-3-butyric acid - NAA -naphthaleneacetic acid - Kn kinetin     

Establishment of regeneration system of callus pathway for Iris sanguinea Donn ex Horn

In Vitro Cellular & Developmental Biology - Plant - Iris is a perennial flowering plant, usually cultivated through seeds or bulbs. However, due to the limitations of traditional reproduction,...