蒙古黄芪组织培养及同源四倍体的诱导与鉴定

通过蒙古黄芪组织培养技术的正交试验，确定了蒙古黄芪最佳繁殖培养基：MS＋BA0．6mg／L＋IAA0．2mg／L＋PP3330．2mg／L；叶作为诱导愈伤组织的外植体优于茎，最适培养基：MS＋BA2．0mg／L＋KT1．0mg／L＋NAA0．5mg／L；最佳生根培养基：1／2MS＋IAA0．5mg／L＋IBA2．0mg／L＋ABT2．0mg／L。建立了蒙古黄芪快速繁殖体系；在无菌条件下利用秋水仙碱诱导蒙古黄芪，获得62个染色体加倍遗传稳定的变异试管苗株系，通过试管苗根尖染色体鉴定，对诱导获得的蒙古黄芪染色体加倍株系进行初步筛选，为进一步获得蒙古黄芪优良品系奠定基础。     

杭白菊同源四倍体的诱导和鉴定

采用正交试验设计方法，进行了杭白菊试管苗快速繁殖技术的优化，同时进行了组织培养条件下杭白菊同源四倍体诱导与鉴定技术的研究。结果表明，诱导愈伤组织的最优外植体为叶片，最适培养基为MS＋KT（激动素）2．0mg／L＋NAA（萘乙酸）0．2mg／L；确定了杭白菊最佳繁殖培养基为MS＋BA（6-苄基嘌呤）0．2mg／L＋IAA（吲哚乙酸）0．05mg／L＋NAA0．2mg／L；最佳生根培养基为1／2MS＋IAA0．2n堰／L＋ABT（生根粉）0．3mg／L。在诱导四倍体的试验中，秋水仙碱浓度对存活率有显著影响，诱导杭白菊同源四倍体的最佳条件是在2％二甲基亚砜和浓度为0．2％的秋水仙碱中浸泡36h，诱导率高达13．33％。通过根尖细胞染色体鉴定，共获得11个同源四倍体株系。     

川麦冬脱病毒和组织培养技术的研究

采用茎尖分生组织培养技术，获得了川麦冬的脱病毒试管苗。通过川麦冬组织培养技术的正交试验及优化筛选，筛选出最佳的培养基组成，用于脱病毒苗的快速繁殖。建立了川麦冬根尖染色体鉴定的最佳条件。结果表明，川麦冬最适繁殖培养基MS＋BA（6-卞基腺嘌呤）2．0mg／L＋NAA（萘乙酸）0．5mg／L；川麦冬最佳诱导愈伤培养基：MS＋BA1．5mg／L＋IAA（吲哚乙酸）0．1mg／L＋2，4-D（2，4-二氯苯氧乙酸）1．0mg／L；通过在培养基中添加不同浓度生长素，得到适合川麦冬的生根培养基：1／2MS＋IAA0．5mg／L＋ABT0．5mg／L。染色体鉴定结果表明：川麦冬的染色体为2n=68。     

兰州百合的快速繁殖及多倍体诱导

为了获得优质品种的兰州百合多倍体,采用鳞片外植体诱导兰州百合(Lilium davidii var.unicolor)试管苗,通过正交实验筛选兰州百合快速繁殖培养基,并对兰州百合鳞茎进行了秋水仙素诱导多倍体的试验研究,还进一步对兰州百合多倍体及二倍体进行了植物抗逆性指标测定。结果表明:兰州百合鳞片能成功诱导出小鳞茎的最佳培养基为MS+PP3332.0 mg/L+BA 1.0 mg/L+NAA 0.1 mg/L+KT 2.0 mg/L,诱导兰州百合多倍体的最佳条件为鳞茎于0.1%秋水仙素溶液浸泡24 h,诱导出的四倍体相比二倍体SOD、POD酶活升高,MDA、脯氨酸含量升高,预示多倍体可能具有较好的抗逆性。     

盾叶薯蓣同源四倍体的诱导和鉴定

采用组织培养技术,运用正交实验设计,进行了盾叶薯蓣试管苗快速繁殖技术的优化和多倍体的诱导与鉴定。结果表明,诱导愈伤组织的最优外植体为叶片,最适培养基为 MS BA2.0 mg/L 2, 4 D0.1 mg/L;丛生芽快速繁殖最适培养基为 MS BA 0.1 mg/L 病毒唑5 mg/L;诱导试管苗生根的最适培养基为MS/2 IBA 0.6 mg/L IAA 0.1 mg/L ABT 2.0 mg/L。在诱导多倍体的试验中,秋水仙碱浓度对存活率有显著影响,诱导多倍体最佳的处理方法为 0.2%秋水仙碱浸泡36 h或 0.3%秋水仙碱浸泡12 h,转接在添加40×10 6秋水仙碱的MS培养基上。     

同源四倍体桔梗组培技术的优化研究

为了建立优良同源四倍体桔梗的快速繁殖和离体保存技术，采用均匀设计和正交设计以添加不同激素的MS培养基为基本培养基进行筛选和优化。结果表明：同源四倍体桔梗最佳丛生芽培养基为MS＋BA（6-苄基嘌呤）0．6mg／L＋NAA（萘乙酸）0．4mg／L＋RTCA（病毒唑）3．0mg／L，丛生芽增殖系数可达到9．13。最佳壮苗培养基为：MS＋BA0．2mg／L＋IAA（吲哚乙酸）0．1mg／L＋PP333（多效唑）0．2mg／L＋RTCA3．0mg／L。最佳生根培养基为：1／2MS＋IAA0．7mg／L＋AC（活性炭）0．2mg／L＋ABT（生根粉）0．2mg／L，生根率可达100％。最适室温离体保存条件为：1／4MS＋蔗糖30mg／L＋甘露醇30mg／L＋PP3331．0mg／L＋BA0．3mg／L＋IAA0．1mg／L＋RTCA3．0mg／L，保存150d时存活率仍可达到59．2％，同时恢复生长快，遗传稳定性好。本研究为优良同源四倍体桔梗的推广应用和优良种质离体保存、减少遗传变异提供了技术依据。     

海南广藿香试管内芽分化与愈伤组织诱导研究

目的选择海南广藿香的最佳芽分化和愈伤组织诱导培养基。方法利用海南广藿香的叶柄、茎尖和带节茎段进行组织培养芽分化试验,并对上述材料进行愈伤组织诱导培养基选择试验。结果在芽分化培养基中,MS＋3-吲哚丁酸（IBA）1．0mg／L＋6-苄基腺嘌呤（6-BA）0．6mg／L培养基能够较好地诱导茎尖和带节茎端长出新芽,诱导率分别为94．3％和94．6％;在愈伤组织诱导培养基中,MS＋萘乙酸（NAA）0．1mg／L＋6-BA2．0mg／L培养基能较好地诱导海南广藿香叶柄和带节茎段产生愈伤组织,诱导率分别为80．0％和100％。结论海南广藿香的最佳芽分化和愈伤组织诱导培养基分别为Ms＋IBA1．0mg／L＋6-BA0．6mg／L和Ms＋NAA0．1mg／L＋6-BA2．0mg／L。     

细辛组织培养技术研究

目的研究细辛组织培养技术,优化细辛组织培养体系。方法以野生细辛越冬芽为外植体,在不同激素浓度配比的培养基上进行初代培养、不定芽增殖及生根培养,并进行生根苗的移栽。结果细辛组织培养过程中适宜的培养基为：（1）初代培养基,MS＋6-BA1．0mg／L＋NAA0．1rag／L;（2）不定芽增殖培养基．MS＋6-BA1．5rag／L＋NAA0．2mg／L;（3）生根培养基,1／2MS＋IBA1．0mg／L。结论优化的培养体系适宜细辛的组织培养,可在短期内获得大量优质种苗。     

白花除虫菊组织培养研究

通过除虫菊组织培养技术的正交试验及优化筛选，建立了除虫菊最适繁殖培养基：MS＋BA0．3mg／L＋NAA0．2mg／L；通过在培养基中添加不同浓度生长素，得到适合除虫菊的生根培养基：1／2MS＋IAA0．2mg／L＋ABT0．1mg／L。建立了除虫菊根尖染色体鉴定的最佳条件。染色体鉴定结果表明：白花除虫菊的染色体为2n=18。为除虫菊的种质保存和优良品种的选育工作奠定了基础。     

怀地黄分生组织培养脱病毒及快速繁殖技术的研究

采用茎尖分生组织培养技术,获得了怀地黄的脱病毒试管苗.通过基本培养基和激素配比试验,筛选出最佳的培养基组成,用于脱病毒苗的快速繁殖.建立了怀地黄根尖染色体鉴定的最佳条件.结果表明:诱导丛生芽的最适培养基为:MS 6-卞基腺嘌呤(6-BA)0.5 mg/L 萘乙酸(NAA)0.1 mg/L 6-糠氨基嘌呤(KT)0.05 mg/L,技术上达到了快速繁殖规模生产的要求.诱导试管苗生根的最适培养基为:1/2MS 吲哚乙酸(IAA)0.05 mg/L.用电子显微镜进行了反复的病毒检测,证明脱病毒彻底,脱毒试管苗将作为今后提供无病毒优质种苗的种源.染色体鉴定结果表明:怀地黄的染色体为2n=2X=56     

Micropropagation and in vitro flowering of Rauvolfia tetraphylla; a potent source of anti-hypertension drugs

A simple protocol for in vitro mass multiplication of Rauvolfia tetraphylla (Apocynaceae) has been developed. The endophytic microflora was controlled by adopting integrated measures. Multiple shoot development was achieved on MS + Kin (0.1-0.2 mg/l) + BAP (0.4-0.5 mg/l) media. Rooting from in vitro shoots occurred on NAA containing media. In vitro flowering was induced in shoot multiplication media.     

胀果甘草愈伤组织诱导培养

试验以甘草的子叶和下胚轴为外植体，分别接种在不同激素组合的MS培养基上，黑暗条件下培养，诱导愈伤组织。结果表明：诱导甘草子叶愈伤组织的最适培养基为MS 2，4-D(1mg／L) BA(1mg／L)，其愈伤组织诱导率达100％，而且其愈伤组织鲜重与接种外植体重的比值达到15．37倍；诱导甘草下胚轴愈伤组织的最佳培养基为MS 2，4-D(0．5mg／L)和MS 2，4-D(0．5) NAA(0．5)，通过在这两种培养基上进行愈伤组织的增殖培养，也得到了十分可观的增殖效果，其相对生长量分别达到了381．52％和498．80％。     

地丁草组织培养及无性系建立的研究

目的:为满足人们栽培对种苗的需要,并保护野生资源.方法:以地丁革不同外植体为材料,进行了愈伤组织诱导与分化、不定芽生根和试管苗生根继代培养研究,建立起地丁革的无性系.结果:MS+ NH4H2PO420 mg·L-1+BA0.2 mg·L-1+IBA0.1mg·L-1+ NAA 0.8～1.0mg·L-1是愈伤组织继代扩...     

Cultures ofGinkgo biloba,effect of nutritional and hormonal factors on the growth of cultured cells derived fromGinkgo biloba

Calli and suspension cultures were obtained following inoculation of the explant from leaves ofGinkgo biloba L. on the supplemented MS basal medium. The obtained calli and suspension cultured cells were able to produce detectable amounts of ginkgolides which are known as natural specific PAF antagonists. The production of ginkgolides in the calli and suspension cultured cells were identified using GC/MS, GC and HPLC with authentic compounds. Since the production of ginkglides A and B in the calli and suspension cultured cells had been confirmed, effects of types and concentration of plant growth regulators, media and illumination on the induction and growth of the callus were studied. The concentrations of growth regulators for optimal callus induction were 1.0 to 2.0 mg/L for NAA and 0.1 mg/L for kinetin. The growth of the callus seemed to be more stimulated with the combination of NAA and kinetin than NAA and BA with illumination at all concentration ranges of 1.0 to 4.0 mg/L for NAA and 0.1 to 1.0 mg/L for kinetin or BA studied. Among 8 different media used, the induction rate of callus on Anderson, Eriksson, and Schenk and Hildebrant at 4 weeks after the innoculation was almost the same as that of MS. However, callus was rarely induced on Heller or White medium. Suspension cultures were easily initiated with 3 g of callus (fresh weight) derived from ginkgo leaves on supplemented MS medium. A typical growth curve of suspension cultured cells could be obtained by measuring the fresh weight of the suspension cultured cells at every 3 days. To improve the growth of suspension cultured cells of ginkgo, effects of concentrations of NAA, sucrose, phosphate ions and molar ratio of NH₄ ⁺ to NO₃ ^? ions in the culture medium were studied. The maximum growth of the cells was achieved when the culture medium contained 1.0 mg/L of NAA, 30 g/L sucrose, 1.75 mM phosphate ions and 1∶5 molar ratio of NH₄ ⁺ to NO₃ ^? ions.     

Shoot Tip Culture of Arnica montana for Micropropagation

Multiple shoots were regenerated from shoot tips of ARNICA MONTANA on MS and B5 media supplemented with BA (1 mg/l) and NAA (0.1 mg/l). Sections of 1-2 mm in length cultured from IN VITRO germinated seedlings regenerated 7.7 (mean) shoots on the MS medium, whereas sections cultured from greenhouse plants regenerated 9.0 (mean) shoots on the B5 medium within 6 weeks. Subsequent subcultures of shoots on the same media but without NAA resulted in similar or lower multiplication rates (1.6 to 3.1 in 3 weeks). Shoot development was promoted, whereas shoot initiation was simultaneously inhibited by the addition of activated charcoal to the media. Rooting was induced by culturing shoots from seedling as well as from greenhouse plant shoot tips on MS or B5 medium supplemented with NAA. The plantlets were transplanted into soil and grown successfully under greenhouse and field conditions.