神农香菊丛生芽的诱导及植株再生

以神农香菊茎段为外植体,研究植物生长调节剂及活性炭(AC)对神农香菊丛生芽诱导的影响,以建立神农香菊丛生芽诱导及植株再生的高频再生体系。结果表明,丛生芽诱导的最适培养基为MS+2.0 mg/L 6-BA+0.05 mg/L NAA,最适增殖培养基为MS+2.0 mg/L 6-BA+0.05 mg/L NAA+0.05 g/L AC,增殖系数为45.3,添加AC可有效促进玻璃化苗恢复;生根诱导的最适培养基为不添加任何生长调节剂的1/2MS,生根率高达100%;将生长良好的再生植株进行移栽,存活率可达100%。     

油葵子叶外植体不定芽再生体系的建立

对油葵子叶外植体在含有不同激素浓度的培养基上进行了诱导、分化及成苗的研究,建立起一套完整的再生体系.在MS添加IAA 1mg/L和6-BA 4mg/L的培养基上诱导的愈伤质量较好,在MS添加IAA 0.01mg/L和6-BA 0.04mg/L的分化培养基上能够实现较高的不定芽再生.基因型间再生能力差异明显,8份材料中以康地6R为最佳.不定芽在1/2MS培养基上能健康成苗.     

蜻蜓凤梨叶片组织培养植株再生的研究

以蜻蜓凤梨试管苗叶片为外植体进行离体再生研究。结果表明，叶片诱导直接分化不定芽的最佳培养基是MS＋BA 5.0 mg/L＋IBA 1.5 mg/L，分化率高达80%。增殖培养基为MS＋BA 0.5 mg/L＋NAA 0.2 mg/L。壮苗培养基为MS＋NAA 2.0 mg/L。生根培养基为MS＋NAA 2.0 mg/L＋AC 0.5 g/L，生根率达100%。     

花生幼叶芽诱导和植株再生研究

从萌发9－10d的花生幼嫩叶片上切取中段作外植体，接种MS＋BA 3mg/L NAA 0.8mg/L AgNO3 2mg/L诱芽培养基，12－14d后产生丛生芽点，芽诱导率达78．9％，平均每外植体产生9个丛生芽。4周后转至MS BA 3mg/L AgNO3 2mg/L诱导芽伸长，诱导生根后获得再生植株。     

Morphogenic Response of Leaf and Petiole Explants of Broccoli Using Thidiazuron

Broccoli (Brassica oleracea var. italica) is an important nutritionally rich vegetable cole crop grown in the world. Environmental stress, pests, and diseases cause enormous yield losses because of a limited gene pool. Genetic manipulation is becoming an important method for broccoli improvement. The objective of present study was to evaluate the potency of thidiazuron (TDZ) as a plant growth regulator in evoking morphogenic responses in leaf and petiole explants of broccoli. An efficient, reproducible, and high frequency plant regeneration protocol has been standardized in broccoli cv. Solan green head. Leaf and petiole explants were cultured on Murashige-Skoog (MS) medium, supplemented with a wide range of TDZ concentrations. The following treatments were designed for efficient in vitro shoot regeneration: TDZ alone, TDZ with adenine, TDZ with naphthalene acetic acid (NAA), and TDZ with indole acetic acid (IAA). Among the 36 combinations of growth regulators used, the highest percentage of leaf explants producing shoot (89.25%) was recorded on MS medium containing 1.0 μM TDZ and 0.107 μM NAA. The multiple shoot regeneration response of petiole explant producing shoots (91.55%) was obtained on MS medium containing 2.0 μM TDZ and 0.107 μM NAA. Shoot multiplication and elongation were obtained on the same medium. For root regeneration in in vitro regenerated shoots, different concentrations of NAA were applied. High frequency (100%) root regeneration response with healthy and vigorous roots was observed on MS medium supplemented with 0.54 μM NAA. The regenerated plantlets with well-developed shoots and root system were transferred to pots containing cocopeat and successfully acclimatized. We recommend 1.0 μM TDZ with 0.107 μM NAA and 2.0 μM TDZ and 0.107 μM NAA combinations for adventitious shoot regeneration from leaf and petiole explants in broccoli cv. Solan green head respectively. This is the first report on high frequency organogenesis from leaf and petiole explants of broccoli cv. Solan green head using thidiazuron.     

Large scale in vitro propagation of Stevia rebaudiana (bert) for commercial application: Pharmaceutically important and antidiabetic medicinal herb

Stevia rebaudiana is a valuable medicinal plant species and it is being used for the treatment of diabetes. Currently, there is a high demand for raw material of this medicinal herb due to ever increasing diabetes disorder among the population. In order to meet the increased demand an efficient in vitro propagation of S. rebaudiana was established. Nodal explants collected from the field were cultured on MS basal medium fortified with different concentrations of BAP (0.5-3.0 mg/l) and KIN (0.5-3.0 mg/l) individually for shoot bud induction. In vitro derived nodal buds were cultured on MS medium supplemented with different concentrations (0.5-3.0 mg/l) of BAP and KIN for multiple shoot bud regeneration. In the second experiment, in vitro derived buds were placed on MS medium supplemented with different concentrations of BAP (0.5-3.0 mg/l) in combination with 0.5 mg/l IAA or IBA or NAA for shoot bud multiplication. The highest frequency (94.50%) of multiple shoot regeneration with maximum number of shoots (15.69 shoots/explant) was noticed on MS medium supplemented with 1.0 mg/l BAP. For large scale plant production, in vitro derived nodal bud explants were cultured on MS medium fortified with 1.0 mg/l BAP, in which about 123 shoots/explant were obtained after three subcultures on the same media composition. Elongated shoots (>2 cm) dissected out from the in vitro proliferated shoot clumps were cultured on half-strength MS medium containing different concentrations of NAA (0.1-0.5 mg/l) and/or MS medium fortified with various concentrations (0.5-2.0 mg/l) of auxins (NAA, IAA and IBA) for root induction. Highest frequency of rooting (96%) was noticed on half-strength MS medium augmented with 0.4 mg/l NAA. The rooted plantlets were successfully transferred into plastic cups containing sand and soil in the ratio of 1:2 and subsequently established in the greenhouse. The present in vitro propagation protocol would facilitate an alternative method for rapid and large-scale production of this important antidiabetic medicinal plant.     

Proline and Glutamine Improve in vitro Callus Induction and Subsequent Shooting in Rice

This study was conducted to evaluate the effects of proline and glutamine on in vitro callus induction and subsequent regeneration and to develop a reproducible and highly efficient plant regeneration protocol in four rice genotypes, viz. Pawana, Jaya, Indrayani and Ambemohar. Considerable variation in response to plant growth regulators and amino acid supplements used was observed in all the four genotypes. Medium supplemented with proline and glutamine was shown to be superior to medium without proline and glutamine. The best callusing from mature embryo was observed on Murashige and Skoog(MS) medium supplemented with 2.0 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D), 500 mg/L proline and 500 mg/L glutamine. Shoot induction was higher in the callus obtained from medium supplemented with 500 mg/L proline and 500 mg/L glutamine. The highest shoot regeneration frequency(83.2%) was observed on MS medium with 2.0 mg/L benzylaminopurine, 0.5 mg/L 1-naphthaleneacetic acid, 500 mg/L proline, and 500 mg/L glutamine in the callus obtained from MS medium supplemented with 2.0 mg/L 2,4-D, 500 mg/L proline and 500 mg/L glutamine. Among the four genotypes, Pawana has the highest regeneration efficiency(83.2%), whereas the regeneration efficiency of the rest three rice genotypes was in the range of 32.0% to 72.3%. This optimized regeneration protocol can be efficiently used for Agrobacterium mediated genetic transformation in rice.     

培养条件对甘蓝型黄籽油菜下胚轴的再生影响

以甘蓝型黄籽油菜GH01的下胚轴为材料，研究影响外植体芽再生的因素。试验结果表明，苗龄、预培养基的激素浓度和预培养时间、分化培养基等对芽再生频率均有较大影响。8d苗龄的下胚轴置MS 1．5mg／L2，4-D 0．1mg／L6-BA培养基预培养4d后，转分化培养基MS 4．0mg／L6-BA 0．05mg／LNAA 5．0mg／LAgNO3 0．6mg,／LGA3培养，可获得较高的芽再生频率。添加5mg／LAgNO3处理可防止外植体褐化并有利于芽分化；0．6mg／L GA3处理可促进胚状体形成，提高芽再生频率。GH01最高芽再生频率为40．50％。     

Regeneration ofSolanum tuberosum cv. Katahdin from leaf expiantsin vitro

A previously published medium has been found to be effective for plant regeneration from leaf expiants ofSolanum tuberosum cv. Katahdin. A number of combinations of thidiazuron and naphthaleneacetic acid, along with a medium previously published for shoot regeneration from protoplast-derived calli ofSolanum phureja, which contained zeatin, indoleacetic acid and gibberellic acid (ZIG), were tested for callus induction, and all calli were transferred to ZIG for shoot induction. We conclude that leaf expiants cultured on ZIG medium at all stages of culture were the most effective at shoot production. A mean of over six shoots were produced per expiant after four months in culture.     

水田七的组织培养和植株再生

用水田七成熟种子为材料进行无菌播种,得到无菌苗再用幼叶、叶柄为材料进行组织培养和植株再生。经试验得出各阶段适宜的培养基分别为：(1)诱导愈伤组织：MS＋2.0 mg/L TDZ;(2)诱导芽分化：MS＋1.5 mg/L 6-BA＋0.5 mg/L NAA;(3)生根培养：1/2MS＋0.5 mg /L IBA＋0.1 mg/L NAA。     

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.     

In vitro regeneration from petiole explants of non-toxic Jatropha curcas

Jatropha curcas, a multipurpose shrub has acquired significant economic potential as biodiesel plant. The seeds or pressed cake is toxic due to the presence of toxic substances and is not useful as food/fodder despite having the best protein composition. A simple, efficient, and reproducible method for plant regeneration through direct organogenesis from petiole explants of non-toxic J. curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). The best induction of shoot buds (57.61%), and number of shoot buds (4.98) per explant were obtained when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 μM TDZ. The Induced shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM 6-benzyl aminopurine (BA), and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation and subsequent elongation was achieved on MS medium supplemented with 2.25 μM BA and 8.5 μM IAA. The elongated shoots could be rooted on half-strength MS medium with 15 μM IBA, 11.4 μM IAA and 5.5 μM NAA with more than 90% survival rate.     

Optimizing regeneration condition in chickpea (Cicer arietinum L.)

In this study, multiple shoot induction and whole plant regeneration from decapitated embryo axes of three chick peal genotypes including MCC252, MCC283 and MCC505 were evaluated on modified Murashige and Skoog's medium (MMS) which, its vitamins were replaced by vitamins of B5 medium, supplemented with varied concentration of thidiazuron (0.1, 0.2 mg L(-1)) or 6-benzylaminopurin (1,2 mg L(-1)) or zeatin (1, 2 mg L(-1)) treatments. BAP was found to be the most effective cytokinin in normal multiple shoot induction. Shoots were elongated on growth regulator-free medium and then rooted on two media containing 1/4 MMS salts and B5 vitamins + 3% sucrose + 0.8% agar with indol-3-butyric acid (0.4 or 1 mg L(-1)). The highest rooting frequency resulted in a medium including 0.4 mg L(-1) IBA. It was found that different shoot induction media also positively affected rooting, where a medium with 2 mg L(-1) BAP in MCC252/MCC505 and a medium with 2 mg L(-1) zeatin in MCC283 were the best media in shoot induction that produced high frequency, thick spread roots. Plantlets were preliminary acclimatized in liquid medium (1/4 MMS salts and B5 vitamins + 3% sucrose + 0.4 mg L(-1) IBA) for 7 to 14 days, then transferred to pots filled by cocopit: perlite (1:1) and kept in a growth chamber until their shoots and roots were well developed. This resulted in more than 70% survival rate.     

High-efficiency regnerationin vitro from potato petioles with intact leaflets

The shoot/plantlet regenerationin vitro of seven potato (Solanum tuberosum L.) cultivars from petioles with intact leaflets was assessed using six treatment combinations-a basal medium with or without silver thiosul-phate (STS) or thidiazuron (TDZ) at two concentrations (2 or 0.5 mg/l) of the indoleacetic acid (IAA). The basal medium consisted of Murashige and Skoog (MS) salts and vitamins supplemented with 3 mg/1 6-benzylaminopurine, and 1 mg/1 gibberellic acid, 30 g/l sucrose, and 7.0 g/l PHYTOAGAR. Two full sets repeats and one partial set repeat of independent experiments were conducted and all produced similar results. Silver thiosulphate decreased the regeneration frequency and number of shoots per callus. No significant changes were observed with thidiazuron. Regeneration rates of (100% ) with up to 20 shoots/plantlets per callus were achieved at 2 mg/1 IAA with cultivars Désirée, Kennebec, Niska, and Lenape. These cultivars still showed high regeneration rates (87%–98% ) on media with 0.5 mg/1 IAA, and good regeneration rates were also achieved by the other three cultivars (48%, 94%, and 50% for Chieftain, Russet Burbank, and Shepody, respectively). Even with the single medium protocol (0.5% IAA without thiosulphate or thidiazuron), Désirée, Lenape, and Niska exhibited a regeneration rate of 98%. The use of petiole-with-leaflet explants could be ideal for the regeneration step of potato genetic transformation protocol because of their high regeneration efficiency and their small cut surface area forAgrobacterium elimination after co-incubation.     

Sprouted Sorghum Extract Elicits Coleoptile Emergence,Enhances Shoot and Root Acclimatization,and Maintains Genetic Fidelity in indica Rice

The high growth-stimulating effect of plant extract has urged the plant biotechnologists to use natural supplements in the culture media instead of synthetic phytohormones. We advocated the effect of sprouted sorghum extract(SSE) on emergence, in vitro acclimatization, and genetic fidelity in coleoptile derived callus of indica rice variety ADT36. The use of SSE with Murashige Skoog medium efficiently acclimatized the root and shoot apical systems. A higher mat and seminal roots(3.4 g biomass) with an efficient shoot primordium elongation were observed with an increase in the concentration of SSE. Seeds treated with SSE medium showed higher germination and earlier coleoptile maturation about 48 h compared to untreated seeds, and there was a higher expression of e EF-1α with an increase in coleoptile length. B5 medium was effective on inducing embryogenic and nodular callus from 3-day-old coleoptile with 3.0 mg/L 2,4-dichlorophenoxyacetic acid and further proliferated effectively with 0.8 mg/L kinetin with a fresh weight of 180 mg. Highly significant regeneration was observed with combination of 2.5 mg/L 6-benzylamino purine and 3.0 mg/L α-naphthaleneacetic acid. The metabolic and genetic profiles of in vitro and directly cultivated plants were the same, examined through Fourier-transform infrared spectroscopy, random amplified polymorphic DNA(RAPD), inter-simple sequence repeat(ISSR) and R-ISSR(combination of RAPD and ISSR) markers, respectively, and thus confirming the significant efficacy of the SSE incorporated medium. Disarmed T-DNA was transformed to coleoptile derived callus through Agrobacterium tumefaciens LBA4404 and confirmed by GUS assay. The T-DNA integration was confirmed by DNA blot analysis using DNA from transient GUS-expressed explants. Thus, SSE can be used as a natural and organic supplement for organogenesis and efficient acclimatizations of shoot and root apical meristems in regenerated plants.     

紫色甘薯的茎尖培养与脱毒

以紫色甘薯品系为试材,研究6-BA和NAA及基因型对紫色甘薯茎尖培养的影响,并采用硝酸纤维素膜—酶联免疫法(NCM-ELISA)和症状法对再生植株进行病毒学检测。结果表明,培养基中6-BA和NAA浓度和配比对紫色甘薯茎尖培养的愈伤组织、不定根和不定芽的诱导有明显影响。6-BA可促进不定芽的诱导,附加1mg/L6-BA的MS培养基为最佳诱芽培养基。基因型对诱芽率也有一定的影响,其中品系B3和B7的诱芽率可达50%。通过病毒学检测,获得12个紫色甘薯品系的脱毒苗,其平均脱毒率为94.7%。     

大豆子叶节再生中植物生长调节剂浓度及基因型筛选

以黑农35为实验材料,对大豆子叶节再生过程中种子的灭菌方法、萌发培养基中植物生长调节剂的浓度、丛生芽分化、生根培养与驯化等过程中植物生长调节剂的浓度进行研究,确定了大豆组培过程中最适合的种子灭菌方法是氯气灭菌法,确定了萌发培养基中的6-BA浓度为2mg/L,丛生芽分化培养基中6-BA浓度为1.70mg/L,GA3浓度为0.5mg/L时,分化率最高（78.1%）生根培养基中添加0.37mg/ LNAA更适合根的生长。采用此方法对北方37个主要栽培大豆品种进行了基因型筛选,确定了合丰25、黑农35、合丰35、绥农10、绥农14等5个比较适合的大豆基因型。     

播娘蒿高频率再生植株因素的研究

以播娘蒿子叶、子叶柄和下胚轴为外植体，研究不同外植体、激素组合以及硝酸银对播娘蒿再生频率的影响。结果表明：外植体、激素以及硝酸银的影响都非常显著；最佳外植体为下胚轴；最佳激素组合为6－BA（2．0mg/L) NAA(0.5mg/L);附加AgNO3(1.7mg/L)可使下胚轴再生频率提高63．42％。诱导生根的最佳培养基为1／2MS＋NAA（0．5mg/L) IBA(0.5mg/L)。     

苎麻茎尖的组织培养及其诱导植株的再生

对苎麻50号品系的茎尖进行组织培养并诱导植株再生。结果表明:最适茎尖转绿和分化的培养基是1/2MS 6-BA1.0mg/L CH300mg/L的液体培养基,茎尖分化率为100%。最适成苗培养基为MS 6-BA0.5mg/L IAA0.1mg/L CH300mg/L,成苗率是53.3%。扩繁培养基为MS 6-BA0.3￣1.0mg/L,繁殖系数达6.7。适宜生根培养基为1/2MS NAA0.01mg/L。最佳茎尖剥取材料是无菌试管苗,茎尖长以0.3mm带1￣2个叶原基为宜。从茎尖接种培养成苗到移栽需4个月左右。     

甘蓝型冬性和半冬性油菜子叶高效快速芽再生体系的建立

以甘蓝型油菜子叶为外植体,研究了不同激素浓度、组合以及预培养等条件对芽再生的影响.将子叶在含1mg/L 2,4-D和1mg/L BA的MSB5培养基上预培养3d后转入芽分化培养基,可明显提高芽再生频率,再生周期也可缩短三分之一,但延长预培养时间则导致再生频率下降.实验中10个冬性和半冬性油菜品种的平均再生频率达81.9%,其中9个品种的芽再生频率均在75%以上,最高可达98.9%.