多花兰种子无菌萌发及离体快繁研究

以多花兰种子为材料,研究了无机盐浓度、植物生长调节剂和光照条件对多花兰种子非共生萌发的影响,在此基础上,通过研究原球茎增殖和分化、芽苗壮苗和生根的培养基配方及培养条件,建立多花兰组培快繁技术体系.结果表明:多花兰种子萌发培养基为1/6 MS十NAA0.5 mg·L-1+6-BA 2.0 mg· L-1+马铃薯泥50g·L-1+AC 1.0g·L-1,光照度为1.25μmol·m-2·s-1,萌发率63.6％;原球茎增殖继代培养基为1/4 MS+6-BA2.0 mg·L-1+NAA 0.5 mg· L1+AC 1 g·L-1+PE 200 g·L-1,繁殖倍数6.5倍/60 d,芽分化率60.2％;再生芽分化培养基为1/4 MS十6-BA2.0mg·L-1+NAA 0.2 mg· L-1+AC 1 g·L-1+PE 200g· L-1,繁殖倍数4.0倍/60 d,芽分化率85.0％;芽苗壮苗和生根培养基为1/6 MS+6-BA 3.0 mg·L-1+NAA 1.0 mg·L-1+AC 1 g·L-1+蔗糖20g·L-1 +PE 200 g·L-1和1/4 MS+6-BA 2.0 mg·L-1+NAA1.2 mg·L-1+AC1 g·L-1十蔗糖20g·L-1+PE200g· L-1,生根率达100％,生根苗移栽成活率90％.此技术可用于多花兰种苗繁育和种质资源保护.     

黄花蒿组培快繁与种质离体保存的研究

以带侧芽的黄花蒿(Artemisia annua L.)茎段为外植体,以MS为基本培养基,进行组织培养和种质保存研究.结果表明,培养基MS 6-BA 1.0 mg L-1 IBA 0.1 mg L-1、MS 6-BA 0.5 mg L-1 IBA 0.1 mg L-1和MS NAA0.1 nag L-1 IBA 0.5 rng L-1可分别用于黄花蒿的芽诱导、增殖和生根培养,培养20 d的增殖倍数为5.5倍,生根率98.3%.培养基MS CCC 1.0 mg L-1、MS CCC 2.0 nag L-1、MS PP3334.0 mg L-1可用作离体保存,连续保存200 d的存活率分别达72.3%、77.0%、69.2%.活力检测表明,黄花蒿种质经保存后的增殖、生根能力没有下降.因此,可通过诱导腋芽增殖建立黄花蒿快繁体系,及在培养基中添加CCC或PP333拼能使材料长期保存.     

小水榕的组织培养与快速繁殖

1植物名称小水榕(anubias barteri). 2材料类别茎尖、茎段. 3培养条件诱导培养基:(1)MS 6-BA 5.0 mg·L-1(单位下同) KT 5.0;增殖培养基:(2)MS 6-BA1.0,(3)MS 6-BA 2.0,(4)MS 6-BA 3.0,(5)MS 6-BA 4.0,(6)MS 6-BA 5.0;生根培养基:(7)MS NAA 0.3 IBA 0.2.各培养基中均添加20 g·L-1白糖、5 g·L-1琼脂,pH 5.8.培养温度25～30℃,诱导和增殖培养的光照度500 lx,生根培养的光照度1 500～2 000 lx,光照时间12 h·d-1.     

三倍体枇杷的茎尖培养及诱导生根

以三倍体枇杷为材料, 研究了不同消毒方式、MS培养基浓度、植物生长调节剂及浓度配比对茎尖培养及诱导生根的影响。结果表明, 初代培养时, 选择生长饱满、健壮的顶芽及适宜的消毒方式, 外植体剥离长度0.5-0.8 cm, 能显著提高茎尖培养的成活率; MS培养基浓度的变化对外植体的褐化没有明显的影响; 最适茎尖的启动培养基为MS＋1.0 mg·L-1 6-BA＋0.5 mg·L-1 NAA, 成活率高达84.8%; 最适组培苗生根培养基为1/2MS＋0.1 mg·L-1 NAA＋0.01 mg·L-1 IAA＋0.3%活性炭, 生根率达66.7%, 每株平均生根2.83条。该研究结果将为三倍体枇杷再生体系的建立及利用转基因技术对三倍体无籽枇杷进行遗传改良奠定基础。     

白玉兰的组织培养和快速繁殖

1植物名称白玉兰(Magnolia denudata). 2材料类别休眠顶芽. 3培养条件芽诱导增殖培养基:(1)MS 6-BA 0.2mg·L-1(单位下同) NAA 0.05,(2)MS 6-BA0.5 NAA 0.05,(3)MS 6-BA 1.0 NAA 0.05,(4)MS 6-BA 0.2 NAA 0.1,(5)MS 6-BA 0.5 NAA0.1,(6)MS 6-BA 1.0 NAA 0.1,(7)MS 6-BA2.0 NAA 0.05;诱导生根培养基:(8)1/2MS IBA0.2,(9)1/4MS IBA 0.5.以上培养基加0.7%琼脂、100mg·L-1肌醇,蔗糖除生根培养基为20 g·L-1外其余均为30 g·L-1,pH 5.8.培养温度20～28℃,光照度1 500～2500 lx,光照时间9～11 h·d-1.     

樱桃种间杂种成熟胚培养及RAPD鉴定

采用胚培养技术对不同组合欧洲甜樱桃×'中国矮樱桃'的杂种胚进行了离体培养,建立了杂种胚萌发、伸长、增殖及生根培养等技术体系.研究结果表明,樱桃杂种胚萌发的最适培养基为MS+3 mg·L-1 6-BA+0.3 mg·L-1 IBA,伸长和增殖培养基为MS+1.5 mg·L-1 6-BA+0.3 mg·L-1 IBA,最佳生根培养基为1/2 MS+0.8 mg·L-1 IBA.利用RAPD技术对获得的杂种进行鉴定分析,结果表明子代与亲代电泳图谱之间存在差异.本实验获得一批杂种为进一步的遗传育种提供了新种质.     

白豆蔻的组织培养

1植物名称白豆蔻(Amomum kravanh). 2材料类别笋芽. 3培养条件(1)诱导培养基:MS 6-BA 3.0 mg·L-1(单位下同) NAA 0.1;(2)增殖培养基:MS 6-BA5.0 NAA 0.1;(3)生根培养基:MS NAA 0.3.以上培养基均添加蔗糖30 g·L-1、卡拉胶5.6 g·L-1,pH 5.8.培养温度26～27℃,光照度1 200～1 500lx,光照时间10 h·d-1.     

山杜英的离体快速繁殖

1植物名称山杜英(Elaeocarpus sylvestris). 2材料类别带腋芽茎段. 3培养条件(1)诱导分化培养基:MS 6-BA1.0 mg·L-1(单位下同) NAA 0.01;(2)增殖培养基:MS 6-BA 1.0 IBA 0.5;(3)生根培养基:1/2MS IBA 0.5.     

PP333对分蘖洋葱试管苗增殖和生根的影响

以MS 0.1 mg·L-1NAA 0.4 mg·L-1 6-BA为增殖培养基,1/2MS 1.5 mg·L-1IBA 0.01 mg·L-1NAA为生根培养基,添加不同浓度PP333的结果表明:增殖培养基中添加1.0 mg·L-1PP333对分蘖洋葱试管苗增殖生长有促进作用,减少超度含水态苗的发生;生根培养基中添加0.1 mg·L-1PP333对分蘖洋葱试管苗生根壮苗有良好效应.     

异源三倍体黄瓜的离体繁殖和鉴定

对甜瓜属双二倍体种 (CucumishytivusChenandKirkbride,2n =4x =38)与栽培黄瓜“北京截头”(C .sativuscv .Bei jingjietou,2n =2x =14 )杂交的种子进行胚拯救。假定为杂种植株的顶芽和腋芽用于离体繁殖的结果表明 :在MS +2 .2mg·L-16 BA和MS +3.0mg·L-1KT +0 .2mg·L-1NAA培养基上诱导不定芽的增殖系数分别为 6 .5 3和 6 .6 3;丛生芽在MS +0 .2mg·L-16 BA的培养基上伸长 ,大约 10d后取整齐一致的芽用于生根 ,在 1/ 2MS +0 .2mg·L-1IBA上生根率达91.7%。田间组培苗的单性结实能力比“双亲”更强 ;果刺黑色 ,与C .hytivus相同 ;叶深绿色 ,不同于双二倍体特有的淡黄绿色而与“北京截头”一致 ;花粉粒可染率不到 10 % ,根尖体细胞染色体鉴定为为 2n =3x =2 6 ,从而确定其为异源三倍体黄瓜。     

药蒲公英再生体系的建立和优化

通过愈伤组织诱导和直接不定芽再生途径 ,建立了药蒲公英的快速高效再生系统。叶片外植体在含0.2mg LIAA和1mg LTDZ的MS培养基中培养 2周后便产生大量的丛生芽 ,在含有 0.5mg/L 2 ,4D和2mg/L 6BA的MS培养基中培养 30d后 ,形成明显的愈伤组织 ,愈伤组织块在含10mg/L 6BA的MS培养基中成功再生。对 9株再生植株进行RAPD检测表明 ,部分植株在DNA水平上发生了变异。与对照相比 ,再生植株的主要抗氧化成分无明显变化 ,保证了有效成分的稳定。     

Thidiazuron stimulates adventitious shoot regeneration in different safflower explants

Adventitious shoot regeneration from root, hypocotyl, cotyledon and primary leaf explants of safflower (Carthamus tinctorius L.) was studied. Shoot regeneration was promoted by benzyladenine (BA) + naphthaleneacetic acid (NAA), BA + indole-3-butyric acid (IBA), kinetin + NAA and thidiazuron (TDZ) + NAA incorporated in Murashige and Skoog (MS) basal medium. High frequency of shoot regeneration and high number of shoots per regenerating explant were obtained on a wide range of TDZ + NAA combinations. Proliferated shoots were elongated in MS + 0.5 mg dm⁻³ kinetin and well-developed shoots were rooted in half strength MS + 0.5 mg dm⁻³ NAA. Rooted shoots were successfully acclimatized and established in soil.     

苹果矮化砧木‘JM7’的组织培养及其离体叶片不定梢再生

以苹果优良矮化砧木‘JM7’ （Malus prunifolia×M. pumila ‘Malling 9’）为试材, 研究了基本培养基对试管苗增殖生长的影响、蔗糖浓度对试管苗生根的影响及基本培养基、细胞分裂素种类和浓度对离体叶片不定梢再生的影响。结果表明： 基本培养基MS比QL显著提高增殖梢数, 但QL比MS更有利于获得健壮生长的绿苗。3%蔗糖浓度比2%的不定根发生速度快。叶片不定梢再生最适宜的基本培养基是QL。在QL培养基上, 6-BA和TDZ对离体叶片不定梢再生率的影响无显著差异, 但6-BA诱导产生的不定芽在不定梢诱导培养基上可直接伸长生长形成不定梢, 而TDZ诱导产生的不定芽需转移到不加TDZ而加低浓度6-BA的培养基上形成伸长生长的不定梢。     

生长调节物质对草莓叶片再生不定芽的影响

MS培养基中添加3.0 mg·L-16-BA和0.1 mg·L-12,4-D的草莓"达斯莱克特"叶片再生频率最高可达94%.2,4-D诱导"达斯莱克特"叶片不定芽的能力明显优于IAA、IBA."童子1号"则以IAA的效果较好.TDz可提高"童子1号"叶片再生频率达80%,但对"达斯莱克特"的诱导效果不及6-BA.另外,KT与6-BA配合诱导不定芽优于单独使用KT.     

Pretreatment in thidiazuron improves the in vitro shoot induction from leaves in Curculigo orchioides Gaertn., an endangered medicinal plant

Leaf regeneration via direct induction of adventitious shoots obtained from an endangered medicinal plant, Curculigo orchioides Gaertn. by pretreating with thidiazuron. C. orchioides is an endangered medicinal herb belonging to the family Hypoxidaceae. Direct inoculation of leaf pieces on MS medium supplemented with various concentrations of BAP (2–8 μM) or TDZ (2–8 μM) alone or in combination with NAA (0.5 and 1.0 μM) produced low shoot induction both in terms of % response and number of shoots per explant. Hence, leaf explants were pretreated with 15, 25 or 50 μM thidiazuron (TDZ), for 6, 24 or 48 h with the aim of improving shoot regeneration from cultured explants. After pretreatment, explants were transferred to an agar solidified MS medium that was supplemented with BAP (4 μM), TDZ (6 μM), BAP (4 μM) + NAA (1.0 μM), TDZ (6 μM) + NAA (0.5 μM). Control explants were incubated directly on the medium without any pretreatment. The pretreatment of explants with 15 μM TDZ for 24 h significantly promoted the formation of adventitious shoots and the maximum response was observed on MS medium supplemented with 6 μM TDZ. In this medium, 96 % cultures responded with an average number of 16.2 adventitious shoots per explant. The percentage of leaf explants producing shoots and the average number of shoots per explant were significantly improved when TDZ pretreated leaves were cultured onto MS medium supplemented with BAP or TDZ alone or in combination with NAA. The rooted plantlets were successfully transplanted to soil with 90% success. The present investigation indicated the stimulatory role of TDZ pretreatment in regulating shoot regeneration from leaf explants of C. orchioides.     

In vitro multiplication of Nothapodites foetida (Wight.) Sleumer through seedling explant cultures

In vitro multiplication of Nothapodites foetida (Wight.) Sleumer was achieved using axenic seedling explant cultures. Isolated nodes (1.0-1.2 cm) and shoot tips (1.0-1.5 cm) cultured in Murashige and Skoog's agar medium containing varying concentrations of TDZ, BA and combinations of 2iP and GA3. Single shoot (0.8-1.2 cm) was regenerated in each culture after 6 weeks. Axillary shoots were then excised and recultured for 8 weeks in medium containing TDZ (0.05 mgL-1) which formed shoots (about 4 in no.; 2 cm) from the basal node. Axillary branches (2) which formed on 60% of these shoots after 10-12 weeks of culture were separated and recultured in the same medium for 8 weeks. Three shoots (0.8-1.0 cm) per culture were regenerated. Shoots of 0.8-1.8 cm length were subcultured on a low cytokinin (0.01 mgL-1 TDZ) regime to induce shoot elongation (2.0-3.5 cm) in 4 weeks. Shoot cuttings were rooted (60%) in the medium containing IBA (1.5 mgL-1). Rooted plantlets established in pots (90%) after hardening resumed normal growth in 3 months.     

Thidiazuron stimulates shoot regeneration of sugarcane embryogenic callus

Summary Efficient shoot regeneration of sugarcane (Saccharum spp. hybrid cv. CP84-1198) from embryogenic callus cultures has been obtained using thidiazuron (TDZ). Callus was placed on modified Murashige and Skoog (MS) medium containing 2.3 μM 2,4-dichlorophenoxyacetic acid (2,4-D), or 9.3 μM kinetin and 22.3 μM naphthaleneacetic acid (NAA) and compared with the same MS medium supplemented with 0.5, 1.0, 2.5, 5.0 or 10.0 μMTDZ, A11 TDZ treatments resulted in faster shoot regeneration than the kinetin/NAA treatment, and more shoot production than either the 2,4-D or kinetin/NAA treatments. Maximum response, as determined by total number of shoots (26 per explant) and number of shoots greater than 1 cm (4 per explant) 4 wk after initiation, was obtained with 1.0 μM TDZ. The shoots rooted efficiently on MS medium supplemented with 19.7 μM indole-3-butyric acid (IBA). These results indicate that TDZ effectively stimulates sugarcane plant regeneration from embryogenic callus, and may be suitable to use in genetic transformation studies to enhance regeneration of transgenic plants.     

菊花叶盘片转基因再生体系的优化选择

以菊花(Dendranthema morifolium)无菌苗叶片为外植体,在芽诱导培养基上直接诱导植株再生,优化选择菊花转基因再生体系。筛选出了能直接诱导芽再生的培养基(MS+6-BA3 mg.L-1+NAA 1 mg.L-1)。茎段在有IBA和NAA的1/2MS培养基上诱导生根。生根的小苗在温室里生长良好。     

欧美杂种山杨愈伤组织再生系统的建立

为解决欧美杂种山杨成年树种快速繁殖的问题,并为研究原生质体分离、遗传转化和体细胞变异打下基础,我们建立了愈伤组织再生系统.其愈伤组织诱导最适培养基为WPM 6-BA 1.0 mg·L-1 NAA 0.1 mg.L-1、WPM 6-BA 1.0 mg·L-1,分化培养基为wPM 6-BA 03～1.0 mg.L-1,愈伤组织诱导率和分化率分别为90.67%和92%.     

采用俄罗斯橄榄叶片和茎段诱导产生不定芽的高效再生方法

俄罗斯橄榄(Elaeagnus angustifolia L.)是一种具有很重要药用价值和生态意义的植物。以俄罗斯橄榄一年生幼苗的叶片和茎段为实验材料,探讨了细胞分裂素类(6-BA和Zt)和生长素类(NAA和IBA)两类激素不同组合以及不同配比对植株再生的影响,最后建立了一个高效的俄罗斯橄榄再生方法。结果表明,MS 培养基+ 0.5 mg/L 6-BA +0.2 mg/L NAA更适合叶片的再生,平均每个外植体能产生多达4.3个不定芽;而在MS培养基 + 1.0 mg/L Zt +0.5 mg/L NAA的条件下,茎段外植体再生出来的不定芽最多可以达到平均3.6个;再生芽在含有0.5 mg/L NAA的1/2 MS培养基上生根率达到100%。体外再生苗移栽到装有灭菌混合土(土∶泥炭∶沙子=1∶1∶1)的花盆中锻炼驯化,最后有77%的再生植株存活下来。此结果不仅对俄罗斯橄榄种质资源保护有重要的促进作用,另外也为其将来的遗传转化奠定了基础。