植物生长延缓剂MET在水稻组织和细胞培养中的作用

目前植物试管苗生长尚不正常,普遍存在着再生苗生长细弱,移栽成活率低,抗逆性差,在试管中不能长期保存等问题,严重影响试管苗的培养效率和进一步开发应用。目前高效植物生长延缓剂多效唑(MET)的应用,为植物再生苗生长发育调控提供了可能条件,本文就 MET 在水稻组织和细胞培养中的作用和影响进行系统研究,取得很明显的培养效果。方法是将再生绿芽或未成熟胚接种在含 MET2.5mg/L 6-BA 2mg/L NAA0.2-0.     

红根草的组织培养与快速繁殖研究

研究了红根草不同采集季节、外植体类型与消毒效果,不同激素浓度和激素组合与芽及根的诱导,及试管苗的移栽。结果表明,不同外植体类型消毒效果为茎节>茎尖>植株抽茎前的生长点;4~6月份采样最佳;MS+BA0.8~1.6mg/L+NAA0.2mg/L、MS+BA0.2mg/L+NAA0.02mg/L、MS+NAA1.0mg/L分别用于初代、继代、生根培养效果最好;试管苗移栽成活率达90%。     

水稻叶和根的愈伤组织诱导及植株再生

植物名称:水稻(Orgza sativa)品种:寒丰一号。材料类别:幼叶、幼根。培养条件:诱导愈伤组织培养基为MS,附加:(1)2,4-D5.0mg/L(单位下同)+BA1.0;(2)2,4-D4.0+NAA2.0+IAA2.0+BA0.5+ZT0.5+KT0.5;(3)NAA5.0+BA1.0;(4)2,4-D1.0+     

龙芽草的组织培养

植物名称:龙芽草(Agrimonia pilosa)。材料类别:种子发芽后,取其下胚轴、子叶、叶为外植体。培养条件:培养基(1)MS+2,4-D0.5(浓度单位:mg/l,下同);(2)MS+2,4-D1+KT0.25;(3)MS+2,4-D1+BA0.2;(4)MS+NAA0.1+BA2;(5)MS+NAA0.2+BA2;(6)1/2MS+NAA0.1。培养温度24～26℃,每天光照12小时,光照度1500—2000lx。生长与分化情况:     

青紫葛的组织培养

植物名称:青紫葛(Cissus discolor),又名青紫藤或花叶粉藤。培养材料:茎尖、茎节、节间及叶片。培养条件:基本培养基为MS培养基(无机盐减半),蔗糖2％,添加LH0.1mg/L,附加激素含量:(1)2,4-D 0.5mg/L(单位下同) BA0.2;(2)2,4-D 2.0 BA0.2;(3)BA1.0 NAA0.01;(4)DA2 NAA1.0。继代培养基为MS BA2     

彩纹海棠的快速繁殖研究

彩纹海棠叶片培养在MS基本培养基中,研究植物的激素、培养基的物理性质对器官形成的影响及试管苗移栽技术等.试验结果表明细胞分裂素BA促进芽的形成和增殖。BA和NAA配合使用,对叶片形成芽有增效作用。通过继代培养,能繁殖大量无根苗。将无根苗转入含有NAA0.2mg/L或IBA ling/L的1/2MS培养基中,诱导生根形成完整植株。诱导叶片形成芽应采用固体培养基;而液体静置培养则有利于促进芽发育成苗和生根。试管苗移栽获得成功,幼苗生长良好。     

提高石刁柏花药愈伤组织的诱导及植株再生率的研究

本文就提高石刁柏花药愈伤组织诱导率及其器官分化率和植株生根,移栽成活率的有关于主要因子进行研究.1、适宜浓度的2,4-D、NAA、BA 配比的1/2MS(铁盐除外)培养基,可诱导芦笋单核期的花药产生较高百分率的花药愈伤组织,各激素适宜浓度为2,4-D 1.0mg/L、NAA 0.5—2mg/L、BA 0.5—2mg/L。诱导率最高可达78.05%。     

生长激素对油菜苔段分化成苗的影响

在B_5培养基上,研究了不同激素、不同激素浓度和比例,以及不同生长素对油菜(Brassica napus L.刁叶青品种)苔段分化成苗的影响。结果表明:①5～10毫克/升的BA,ZT和KT均可使苔段分化成苗,但BA和ZT效果好,KT效果差。②激素配合使用,有利于苗的分化,其中以BA2毫克/升 ZTl毫克/升效果最好,分化频率高,苗数多,生长正常。③激素和生长素配合使用,对苗的分化也有明显影响。在BA2毫克/升 KT1毫克/升的基础上,附加0.5毫克/升的2,4-D、NAA,或IAA均抑制苗的分化。若同时追加0.5毫克/升的NAA IAA,NAA GA_3,仅分化出大量的根,而未分化成苗。④激素和GA_3配合使用,明显促进苗的分化和生长。⑤在B_5 3％蔗糖的培养条件下,能成功地诱导根的生长。由于从苔段能分化出大量的苗和诱导生根,并形成完整的植株,这对油菜的育种和繁殖均有利用价值。     

金荞麦的组织培养

植物名称:金荞麦(Fagopyrum cymosum)。材料类别:种子无菌苗的下胚轴和子叶柄。培养条件:诱导愈伤组织培养基:(1)MS+2,4-D1mg/l(单位下同)+KT0.25;(2)MS+2,4-D1+BA0.2。分化培养基:(3)MS+NAA0.1+BA2。促进不定芽伸长培养基:(4)MS_0。诱导生根培养基:(5)1/2MS_0;(6)1/2MS+NAA0.1;(7)1/2MS+NAA0.5。培养温度26±2℃,光照度1500—2000 lx,每日照光12小时。     

药用植物金荞麦愈伤组织诱导与植株再生

目前转基因技术已成为植物定向遗传改良的重要手段,而建立稳定高频的离体再生系统是实现遗传转化的基础和前提.本试验以25 ～30 d苗龄的金养麦(Fagopyrum dibotrys)无菌苗叶片、茎节间、叶柄为外植体进行愈伤组织诱导与植株再生研究.结果表明:叶片在MS +2,4-D 4.0 mg/L +6-BA 1.0 mg/L培养基上愈伤组织诱导率达到89％.茎节间在MS +2,4-D 2.0 mg/L +6-BA 2.0 mg/L培养基上愈伤组织诱导率为87％.叶柄在MS +2,4-D 4.0 mg/L +6-BA 2.0 mg/L+ IBA 0.2 mg/L培养基上的最高诱导率仅为54％.愈伤组织分化不定芽的适宜培养基为MS +6- BA2.0 mg/L +TDZ0.2 mg/L +NAA0.2 mg/L;金荞麦不定芽在1/2 MS +NAA 0.5 mg/L的培养基上生根效果最好.组培再生植株经炼苗后移栽到田间成活率达80％以上,且生长表现正常.高频完整再生体系的建立,为金荞麦进一步遗传操作和扩大药材资源奠定了基础.     

甘蓝型油菜与芝麻菜体的细胞杂交

为拓宽油菜育种的基因资源库, 改良油菜品种, 以甘蓝型油菜(Brassica napus)花油3号下胚轴和芝麻菜(Eruca sativa)下胚轴为材料分离制备原生质体; 然后采用PEG-高Ca2+-高pH法进行原生质体融合, 当PEG浓度为35%, 原生质体融合密度为5×105个/mL时, 融合25 min时, 融合率可达18.2%。融合后在培养密度为1×105个/mL时, 以附加1.0 mg/L 2,4-D +0.5 mg/L 6-BA+0.5 mg/L NAA+ 200 mg/L肌醇+300 mg/L水解酪蛋白的改良的KM8p为融合体培养基, 以0.1 mol/L 蔗糖+0.2 mol/L葡萄糖+0.2 mol/L甘露醇作渗透稳定剂进行液体浅层培养, 效果较好, 愈伤组织再生率最高为6.8%。将融合体再生的小愈伤组织转移至培养基(B5无机盐+0.087 mol/L蔗糖+0.2 mg/L 2, 4-D+0.5 mg/L NAA+0.2 mg/L 6-BA+ 0.5% Agar, pH 5.8)上增殖培养, 待愈伤组织长至直径为3~5 mm时, 及时将其转至分化培养基(MS无机盐+0.087 mol/L 蔗糖+0.1 mg/L IAA+0.8 mg/L 6-BA+0.8% Agar, pH 5.8)中诱导不定芽再生, 芽分化率为35.7%。当不定芽长为2~3 cm时, 将其切下转入附加0.5 mg/L IBA+0.2 mg/L 6-BA的1/2MS生根培养基中诱导生根, 14 d左右即可形成再生植株, 生根率可达88%。同时, 以紫外线(60 μW/cm2)照射芝麻菜原生质体, 进行不对称融合, 照射2 min的获得了愈伤组织和再生植株, 照射4 min的只获得愈伤组织, 而照射5 min以上的没有获得愈伤组织, 但其愈伤组织再生、增殖及植株再生均不如对称融合。从细胞学鉴定的21块杂种愈伤组织上再生出16株杂种植株。     

甘蓝型油菜与芝麻菜体的细胞杂交

为拓宽油菜育种的基因资源库, 改良油菜品种, 以甘蓝型油菜(Brassica napus)花油3号下胚轴和芝麻菜(Eruca sativa)下胚轴为材料分离制备原生质体; 然后采用PEG-高Ca2+-高pH法进行原生质体融合, 当PEG浓度为35%, 原生质体融合密度为5×105个/mL时, 融合25 min时, 融合率可达18.2%。融合后在培养密度为1×105个/mL时, 以附加1.0 mg/L 2,4-D +0.5 mg/L 6-BA+0.5 mg/L NAA+ 200 mg/L肌醇+300 mg/L水解酪蛋白的改良的KM8p为融合体培养基, 以0.1 mol/L 蔗糖+0.2 mol/L葡萄糖+0.2 mol/L甘露醇作渗透稳定剂进行液体浅层培养, 效果较好, 愈伤组织再生率最高为6.8%。将融合体再生的小愈伤组织转移至培养基(B5无机盐+0.087 mol/L蔗糖+0.2 mg/L 2, 4-D+0.5 mg/L NAA+0.2 mg/L 6-BA+ 0.5% Agar, pH 5.8)上增殖培养, 待愈伤组织长至直径为3~5 mm时, 及时将其转至分化培养基(MS无机盐+0.087 mol/L 蔗糖+0.1 mg/L IAA+0.8 mg/L 6-BA+0.8% Agar, pH 5.8)中诱导不定芽再生, 芽分化率为35.7%。当不定芽长为2~3 cm时, 将其切下转入附加0.5 mg/L IBA+0.2 mg/L 6-BA的1/2MS生根培养基中诱导生根, 14 d左右即可形成再生植株, 生根率可达88%。同时, 以紫外线(60 μW/cm2)照射芝麻菜原生质体, 进行不对称融合, 照射2 min的获得了愈伤组织和再生植株, 照射4 min的只获得愈伤组织, 而照射5 min以上的没有获得愈伤组织, 但其愈伤组织再生、增殖及植株再生均不如对称融合。从细胞学鉴定的21块杂种愈伤组织上再生出16株杂种植株。     

Regeneration of plantlets from the callus of stem segments of adult plants of Ficus religiosa L.

Stem segments of adult plants of Ficus religiosa L. cultured on MS medium containing 1.0 mg/l 2,4-D produced callus. Shoots were regenerated when the induced calli were transferred to medium supplemented with 0.05 to 2.0 mg/l BAP. Callus derived shoots produced roots and developed into plantlets when transferred to medium supplemented with 1.0 mg/l NAA.Abbreviations MS Murashige and Skoog (1962) - BAP 6-benzylaminopurine - NAA naphthaleneacetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid     

在1/3海水培养基上筛选豆瓣菜耐盐变异体

The responses of stem segments of watercress ( Nasturtium offtcinale R. Br. ) to 6-BA, NAA and 2,4-D were studied. MS medium supplemented with 2.0 mg/L 6-BA, 0.2 mg/L 2,4-D was used for callus initiation and maintainance. MS medium supplemented with 4.0 mg/L 6-BA was suitable for plant regeneration and MS medium without plant hormone supplement was used for rooting and plant propagation. For screening of salt. tolerant calli, stem segments of watercress were plated onto callus initiation medium containing 1/3 natural seawater. Seventeen out of the 325 plated explants produced calli. The growth curves demonstrated that the growth rate of salt-tolerant calli on saline medium almost matched that of the control calli on normal medium. Some of the salt-tolerant calli were transferred to the normal regeneration medium or saline regeneration medium to induce plant regeneration. In the first case, buds and shoots were regenerated in the same way as those of control calli on normal regeneration medium. More than 1 000 regenerated shoots were obtained of which 83 regenerated shoots were cut and transferred to saline MS base medium. At first, all shoot growth was inhibited, but 40 days after the transfer, rapid-growing axillary shoots were observed on 16 of the original shoots but none on the control shoots on saline MS base medium. Moreover, green spots appeared on most calli 10 days after they were transferred to saline medium, however buds appeared only on 5 calli from the 30 transferred calli and at the end only 2 rapid-growing shoots were obtained from two calli. In total, 18 variant lines were obtained through propagation of the salt-tolerant shoots on saline MS base medium. RAPD analysis was performed in 10 of the 18 salt-tolerant variant lines and DNA variation was detected in all the tested variant lines.     

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。     

唐古特大黄组织培养技术的研究

试验选用唐古特大黄(Rheum tunguticum Maxim．ex Regel．)种了萌发的无菌苗及无菌苗子叶、下胚轴、胚根和幼根作为材料,研究唐古特大黄不同外植体的离体培养技术。结果表明,唐古特大黄的无菌苗和无菌苗子叶、下胚轴、胚根和幼根都可以作为离体培养的良好外植体。唐古特大黄的最适分化培养基足：B5 NAA0．1mg／L 6-BA3mg／L;最适乍根培养素是：1／2MS NAA1mg／L 3%蔗糖或1／2MS NAA0．5mg／L 3％蔗糖;愈伤组织诱导培养基是：MS 2,1-D 1mg／L NAA1mg/L 6BA1mg／L。     

鱼腥草组织培养研究

以鱼腥草带侧芽茎段为外植体进行组培快繁研究。结果表明,初代培养以MS+6-BA1.0mg/L+NAA0.05mg/L为最佳,30d侧芽诱导率可达66.7%;MS+6-BA 2.0 mg/L+2,4-D 0.2mg/L+NAA 0.2mg/L对芽苗的继代增殖效果最好,40d芽苗的增殖倍数可达7.4,且芽苗较高;以MS+6-BA3.0mg/L+2,4-D 0.5mg/L+NAA0.1mg/L对芽苗的继代增重效果最好,40d芽苗鲜重为7.358g;以沙和蛭石(1:1)为基质瓶外生根效果最好,30d芽苗生根率可达84%。     

猕猴桃高频直接再生体系的建立

为了建立猕猴桃高频再生体系,以MS为基本培养基,猕猴桃(Actinidia deliciosaQinmei)茎及叶片为外植体,研究了2,4-D、6-BA和NAA在美味猕猴桃愈伤组织形成及分化过程中的作用。方差分析结果表明,6-BA能够显著促进愈伤组织形成,6-BA和NAA可以显著促进愈伤组织形成和分化,而2,4-D抑制愈伤组织形成。附加2.0 mg/L 6-BA、1.0 mg/L NAA和600 mg/L水解酪蛋白的MS培养基是茎段培养的最佳培养基,在该培养基上,以再生的无菌苗为起始材料,一个月时叶圆盘的直接再生频率达到100%,平均每个叶圆盘产生9.33个芽,其中23.21%芽高度超过0.5 cm。     

Plant regeneration from the embryogenic calli of five major Miscanthus species, the non-food biomass crops

With the growing shortage of oil, coal, and other traditional fossil fuels, scientists in various fields have been looking for new fuel sources to solve the energy crisis. The genus Miscanthus is an ideal biofuel crop due to its rapid vegetative growth and its potential for high biomass yields. Plant regeneration through somatic embryogenesis is a viable method to achieve large-scale production of plant biomass. Callus induction from immature inflorescences of five Miscanthus species was performed on two different media, and the relative rates of callus proliferation were calculated. The highest multiplication coefficient, 3.92, was obtained with M. sacchariflorus ssp. lutarioriparius when the concentration of 2,4-dichlorophenoxyacetic acid (2,4-D) in the medium was 4.0 mg/L, and this species also performed the best at the induction phase. The proliferation coefficient for M. floridulus increased to 3.19 when the concentration of 2,4-D was decreased from 4.0 to 2.0 mg/L. When the concentration of 2,4-D was 2.0 mg/L, the proliferation coefficient for M. sinensis was 2.47. In M. sacchariflorus, the proliferation coefficients were 2.89 and 2.49 for the lower and higher concentrations of 2,4-D, respectively. The multiplication coefficient of M. x giganteus was 2.60 on medium containing 4.0 mg/L 2,4-D. Three different regeneration media were tested to induce shoots in vitro. M. floridulus and M. sacchariflorus regenerated shoots at 100% frequency in three different regeneration media. The regeneration rate for M. sacchariflorus ssp. lutarioriparius reached 99.0% on medium containing 4.0 mg/L?N ⁶ -Benzylaminopurine (6-BA) and 0.1 mg/L α-Naphthaleneacetic acid (NAA). The best regeneration rate of M. sinensis was 35.2% using 2.0 mg/L 6-BA and 0.3 mg/L NAA, whereas the regeneration rate of M. x giganteus was 57.4% on medium supplemented with 3.0 mg/L 6-BA and 0.2 mg/L NAA. The in vitro-derived plantlets of all five species had 100% rooting rates on basal MS medium without supplementation. The survival rates of plantlets were above 90% for each species when subsequently grown outdoors.