海巴戟天的离体快速繁殖(简报)

从多果无病虫害的母树中选取的幼嫩侧枝,经表面消毒后切取侧芽并接种到MS基本培养基,附加蔗糖浓度30g/L,BA2mg/L,NAA0.1mg/L初代培养基上,培养40d后,侧芽萌发率达80%以上。增殖培养基与初代培养基相同,以40~50d为一增殖周期,增殖系数达2~3倍。当新芽增殖到足够数量时,切取2~3cm的新芽接种到1/2MS大量元素,全量微量元素,蔗糖浓度为30g/L,IBA1mg/L,活性炭1g/L的生根培养基生根培养基上,生根率高达100%,植株在沙床的移栽成活率均达95%以上。30cm高的植株便可种植到大田。     

银后粗肋草的离体培养和快速繁殖

以银后粗肋草的幼嫩叶片为材料,研究不同植物生长调节剂组合对愈伤组织诱导和分化的影响;以带侧芽的茎段为材料,研究不同接种方式、不同6-BA和NAA组合对丛生芽诱导的影响;以愈伤组织诱导的不定芽和茎段诱导的丛生芽为材料,研究不同植物生长调节剂组合对丛生芽的增殖和生根的影响;最后研究不同移栽基质组合对试管苗移栽成活的影响。结果表明:(1)叶片接种在MS+6-BA 2.5 mg/L+NAA 0.1 mg/L培养基培养中60 d后,愈伤组织诱导率为63.41%,不定芽再生率为26.88%,平均生成芽数为3.80个。(2)带侧芽的茎段水平放置接种在MS+6-BA 2.0 mg/L+NAA 0.1 mg/L培养基上,丛生芽诱导率为91.91%,平均生成芽数为5.29个。(3)将经叶片诱导愈伤组织分化所得的不定芽和经茎段诱导所得的丛生芽切成单株,接种到MS+6-BA 3.0 mg/L+NAA 0.1 mg/L或MS+KT 3.0~4.0 mg/L+NAA 0.1 mg/L增殖培养基中,不定芽和丛生芽增殖和生长良好。(4)将在增殖培养基中长至4 cm以上的小植株转入到1/2 MS+NAA 0.2 mg/L的生根培养基中培养20 d后,生根率达100%,平均根长4.49 cm,每株平均根数6.80条。(5)生根苗移栽到1/2木屑+1/4珍珠岩+1/4菜园土混合基质中,成活率达95.40%。     

蜜糖文心兰的组培快繁技术

以文心兰盆栽品种‘蜜糖'的侧芽为实验材料,研究基本培养基、激素配比、pH值、蔗糖浓度和添加物等对原球茎增殖的影响,探讨文心兰的生根壮苗与炼苗移栽技术.结果表明:(1)文心兰侧芽茎尖在MS+6-BA 4.0mg/L+NAA 1.0mg,L的培养基上能形成原球茎;培养基的pH值为5.8时较有利于原球茎的增殖;最适宜原球茎增殖的培养基为:MS+6-BA 2.0 mg/L+NAA 0.2 mg/L+蔗糖30 g/L+10%香蕉泥.(2)原球茎在MS+6-BA 0.6 mg/L+NAA 0.1 mg,L+蔗糖30 g/L的培养基上能分化出芽,萌芽率可达83.4%;无根苗在1/2 MS+IAA 0.2 mg/L+10%香蕉泥+蔗糖30 g,L的培养基上能形成完整植株,培养35 d后的生根率为92.5%,且假鳞茎饱满,植株健壮,试管苗移栽在水苔上的成活率可达94.6%.     

巨桉组培育苗技术初探

以当年巨桉实生苗顶芽、幼嫩侧芽为繁殖材料,以培养基MS＋6-BA 0.25 mg/L＋IBA 0.3mg/L＋蔗糖35g/L＋卡拉胶6.2 g/L,诱导获得丛芽,在培养基MS＋6-BA 0.25 mg/L＋IBA 0.3 mg/L＋NH4NO3 3 mg/L＋蔗糖35 g/L＋卡拉胶6.2 g/L上继代增殖,增殖倍数2.5以上,生根诱导在MS＋NAA 0.25 mg/L＋IBA 0.3 mg/L＋蔗糖25 g/L＋卡拉胶5.2 g/L的培养基上获得健壮的生根苗,生根诱导率90%以上,移栽苗圃获得成功。     

大豆(Glycine max L.)茎尖的离体培养和繁殖

本文以两个大豆品种为材料,研究了利用茎尖外植体进行离体快繁的可行性.结果表明:以MSB为基本培养基,附加不同浓度的NAA和KT均能促进侧芽增殖.其中,以NAA0.2mg/L、K T0.5mg/L效果最好。侧芽切割后接种于1/2 MS附加NAA0.1mg/L的培养基上可诱导生根,完整植株移栽后极易成活.     

鼓槌石斛组培快繁技术体系研究

以鼓槌石斛八成熟蒴果为外植体,通过对其种子无菌萌发培养、原球茎继代增殖及生根壮苗培养基的筛选,建立鼓槌石斛的组培快繁技术体系。结果表明：1/2 MS＋1.0 mg/L NAA＋8.0%土豆泥＋0.10%活性炭培养基效果较理想,种子萌发率较高,发芽快;最适增殖培养基为3/4 MS+3.0 mg/L 6-BA＋8%香蕉泥,增殖系数可达7.23;最佳生根培养基为3/4 MS＋1.0 mg/L NAA＋0.5 mg/L CA＋8%香蕉泥,平均根数量达7.49条。炼苗移栽到松树皮作为栽培基质的苗床上,1个月以后,成活率均达80%。     

红锥组织培养与快速繁殖的研究

以红锥茎段为外植体, 研究了不同激素对其再生体系建立的影响。结果表明：最适诱导培养基为 BMT＋6-BA 0.3 mg/L＋NAA 0.25 mg/L,诱导率达到了85%;最适增殖培养基为BMT＋6-BA 0.3 mg/L＋NAA 0.5 mg/L,芽增殖系数3.58,且生长健壮;单芽在1/2 BMT＋IBA 1.5 mg/L＋IAA 0.3 mg/L培养基上生根效果好,生根率82%;炼苗成功后移栽成活率达90%以上。     

猫尾射的组织培养

以猫尾射无菌播种苗(去除根部)为外殖体,对其愈伤组织诱导和分化及其不定芽增殖进行研究。结果表明,外殖体在MS＋6-BA 1.0 mg/L＋NAA 0.1 mg/L培养基上,愈伤组织诱导和分化的效果较好,愈伤诱导率为82.0%,分化率为74.5%;经不定芽增殖培养基MS＋6-BA 0.5 mg/L＋NAA 0.05 mg/L培养,不定芽增殖倍数为8.2,平均株高为4.7cm。组培苗在MS＋IBA 0.5 mg/L培养基上生根率达90%。生根苗移栽成活率达84%。     

杜鹃红山茶离体快速繁殖

以杜鹃红山茶实生小苗为试验材料,探讨离体快速繁殖的方法。结果表明:茎段外植体在MS+BA 1 mg/L+IBA 0.01 mg/L+GA3 10 mg/L的培养基上培养60 d,侧芽萌发率高达68%;将这些萌发的侧芽连同原来的茎段外植体移至添加BA 0.2～0.4 mg/L的培养基培养1个月,74%的侧芽能够继续生长并形成丛生芽;再将丛生芽移至1/2 MS+IBA 4 mg/L+0.1%活性炭的培养基上培养1个月,得到大量伸长的芽条;将芽条切离母体后插植于1/2 MS+IBA 8 mg/L的培养基,60 d后生根率达到90%。通过以上过程繁殖得到的小苗质量良好,移栽1个月后的成活率可达90%。     

甜菜组织培养与植株再生体系的建立

本研究以甜菜品种"甘糖7号"叶柄为外植体,通过诱导愈伤组织、愈伤组织分化不定芽、芽生根和移栽等步骤建立一套甜菜组织培养与植株再生体系。结果表明,诱导甜菜叶柄愈伤组织的最佳培养基为MS+4 mg/L 6-BA+0.5 mg/L NAA,诱导率达88.2%;诱导愈伤组织分化不定芽的最佳培养基为MS+4 mg/L 6-BA+0.5 mg/L NAA,分化率仅为9.4%;生根培养基为MS+0.1 mg/L NAA,生根率达33.3%。甜菜再生苗移栽至蛭石中,保温保湿7~10 d后,成活率可达90%以上。本研究结果将对糖料作物甜菜抗逆遗传改良及其基因工程育种具有推动作用。     

山东茶树良种组织培养及繁殖能力的研究

取茶树嫩茎段作外植体进行离体培养 ,以MS作基本培养基 ,经添加不同激素水平及组合的试验 ,得到最佳分化率 83% ,其培养基配方为MS +BA 1 5+ZT 0 5;最佳生根率 80 % ,其培养基配方为 1/ 2MS +IBA0 2 +2 0g糖。选出了最佳移栽基质为蛭石 +壤土 +木屑 ,其配比为 2∶1∶1,移栽成活率达 85%。一个外植体经三次继代培养可繁殖 4 7棵苗。     

根癌农杆菌介导大果西番莲基因转化

以大果西番莲种子无菌苗的子叶和下胚轴为外植体,在添加不同质量浓度BA和IAA的MS系列培养基上进行不定芽的诱导,确定MS+BA2.0mg·L-1+IAA0.1 mg·L-1为最适的不定芽诱导培养基,下胚轴、子叶外植体诱导芽分化率分别为88.33％,90.00％。将诱导出的不定芽接种在添加不同质量浓度IBA和NAA的MS系列培养基上,进行不定根的诱导,确定MS+NAA0.5mg·L-1+IBA0.2mg·L-1为最适的不定根诱导培养基,生根率达92.86％。用三亲交配法将质粒pBICr先转移到E.coli DH5α,然后转移到根癌农杆菌。利用含有NPT-Ⅱ基因和CaMV35S启动子控制下的黄瓜花叶病毒外壳蛋白（CMV-CP）反义基因的表达载体,通过根癌农杆菌介导法将其导入西番莲受体细胞,在质量浓度为50mg·L-1的Kan+选择压力下培养,获得大果西番莲CP转基因再生植株。      

An improved and efficient micropropagation of Eclipta alba through transverse thin cell layer culture and assessment of clonal fidelity using RAPD analysis

An improved and efficient in vitro regeneration system has been developed for Eclipta alba, a medicinally important plant, through transverse thin cell layer culture (tTCL). The transverse section of the nodal segment of field grown plants was used as tTCL explants for plant regeneration. Shoot multiplication from tTCL nodal explants was influenced by BAP and their interaction with Kin or NAA. MS medium containing 13.2 μM BAP and 4.6 μM Kin was most effective for shoot multiplication from tTCL nodal explants. Upon this medium, percent response for shoot proliferation was 100% with an average of 32.6 shoot buds per tTCL nodal explant. Regenerated shoots from tTCL nodal explants were rooted on the growth regulator free MS medium. The rooted plantlets were successfully acclimatized and established in soil with a survival frequency of 90-100%. Random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic fidelity of the micropropagated plants. RAPD profile analysis indicated that micropropagated plants were genetically similar to mother plant.     

In vitro propagation and regeneration of an industrial plant kenaf (Hibiscus cannabinus L.)

The present study report a protocol for the efficient in vitro propagation of kenaf (Hibiscus cannabinus L., an industrial crop having high cellulosic fiber content) on hormone free MS medium using the shoot apex and nodal explants. Shoot tips and nodes were isolated from 15 days old seedlings cultivated on MS medium. Different combinations and concentrations of auxin/cytokinin were used and added to the MS medium to assess the shoot and root induction of theses explants. Several subcultures were drived in order to enhance the multiplication rate. Healthy and well developed in vitro propagated shoots were transferred for acclimatization under greenhouse conditions in pots filled with different substrates (sand + compost or perlite). Our results showed that shoots could elongate and root within 4-6 weeks on MS basal medium without any callus formation. However, addition of growth regulators to the MS medium leaded to a decrease in shoot and root induction rates. Indeed, the highest shoot regeneration frequency (90.5%) was obtained on MS control medium. Elongated shoots were transferred onto the same hormone free MS medium using five subcultures where the multiplication rate reached the highest value (3.66) at the fifth and last step. The in vitro rooted plantlets were acclimatized in greenhouse and successfully transplanted to natural conditions with 70% survival.     

Nursery Growth of Banana (Musa spp.) Plantlets Rooted on Auxin-free and Auxin-supplemented Media

Abstract

This paper describes the effects of auxin added to the culture medium on main and branch root formation of banana (Musa spp.) shoots and growth characters of the plantlet rooted on the medium with and without auxin. Banana shoots cultured in vitro on Murashige and Skoog medium supplemented with 2 μM 1-naphthylacetic acid (NAA), rooted earlier and also had more adventitious roots than those cultured on the medium without NAA. However, the adventitious roots formed on the medium without NAA showed more lateral branching. Plant height and number of leaves per plantlet in in vitro culture were not influenced by the addition of NAA but under nursery conditions, plantlets rooted without NAA showed better growth in terms of days to the appearance of new leaf, plant height and number of leaves per plant. This might be due to the presence of abundant lateral roots. Even though auxins are generally known to promote rooting, NAA inhibited the formation of lateral roots in Banana plants.     

Plant regeneration from cell suspension culture of potato (Solanum tuberosum L.)

In the present study, for callus production leaf and stem segments of potato cultivar White Desiree were cultured on MS medium supplemented with 2,4-D, NAA and Kinetin (callus production medium). Calli then were transferred in the same liquid medium for cell suspension production. In the next step cell suspensions were transferred back to the callus production medium. Finally, calli derived from cell suspension were cultured on 6 different shoot initiation media (S1-S6). However, on S6 medium with combination of GA3 and BAP more than 80% of the calli produced shoot buds and shoots. Fully grown shoots then were rooted and produced whole plants. Chromosome and morphological analysis showed no somaclonal variation among regenerated plants.     

少核‘年桔’的辐射育种研究

用60Co-γ射线4、6、8 kR辐射‘年桔’秋梢芽条,嫁接成活率分别为68.4%、41.3%和16.5%,对照成活率为93.2%。通过连续2 a的果实调查分析,获得少核性状稳定、综合性状优良的突变体4个,平均单果种子含量3.3～6.6粒,极显著低于对照的17.6粒。少核‘年桔’突变体的花粉生活力在45.5%～68.8%,发芽率在11.8%～16.7%,均分别极显著低于对照的97.3%和54.1%,说明少核‘年桔’少核的主要原因是花粉败育。     

铁兰液体振荡培养再生与快速繁殖

以铁兰叶片为外植体,诱导获得脆性愈伤组织,液体振荡进行胚性愈伤组织培养及快速增殖,固体培养诱导胚状体萌发、不定芽生根,最后移栽成活,实现了铁兰的快速繁殖。在优化的培养条件下,铁兰无菌苗叶片愈伤组织诱导率90%以上,振荡后的愈伤组织100%成为胚状体,体积增殖系数可达5.0;90%以上的胚状体可分化为不定芽,95%以上的不定芽可诱导生根,移栽成活率95%以上。研究结果表明成功进行了铁兰的液体振荡培养及快速繁殖。     

农杆菌介导将雪莲凝集素(GNA)基因转入籼稻单倍体微芽的初步研究

 以含有双元载体(携带GNA基因,nptⅡ基因)的根癌农杆菌菌系LBA4404转化籼稻单倍体无性系微芽Hu18,经共培养后在含G418的保存培养基中连续筛选G418抗性芽,抗性芽经生根培养基中壮苗获得单倍体转化植株。PCR分析证明GNA基因已进入到Hu18细胞中,抗虫鉴定表明转基因植株具有白背飞虱抗性。并探明了G418对单倍体微芽的致死浓度和时间,共培养时间对G418抗性芽产生的影响。      

丛生芽—蝴蝶兰无性快速繁殖的新途径

不经过愈伤组织直接诱导蝴蝶兰“丛生芽”是一种变异率低的快繁方法。本文着重研究了诱导“丛生芽”的两个关键环节,即“丛生芽”的增殖及其壮苗的培育。随着BA浓度的提高,“丛生芽”增殖率上升,BA浓度分别为1,5,10,20mg/L培养50天后增殖率分别为189％,307％,475％,523％,但10mg/L培育的苗比20mg/L的壮。添加椰子水可使“丛生芽”的增殖率提高近1倍,而且生势较好。适当增加培养室的光照并加入香蕉和土豆等有机物能促进根系生长,使蝴蝶兰苗更健壮。