几种抗生素对梨叶片愈伤组织和不定梢诱导的影响

研究了氨苄青霉素(Amp)、羧苄青霉素(Carb)、头孢霉素(Cef)和卡那霉素(Km)对梨叶片愈伤组织和不定梢诱导的影响。结果表明,Amp对巴梨叶片分化影响较小,当浓度达500mg/L时,其出愈率和不定梢再生率分别为96.48%和56.17%,与对照差异不显著;Cef浓度在50-300mg/L时对巴梨和身不知叶片再生均无显著影响,只有当Cef浓度高于400mg/L时,巴梨的不定梢再生率才受到显著抑制;巴梨和身不知叶片对Carb的反应不相同,低浓度(≤200mg/L)的Carb对巴梨的出愈率和不定梢再生率影响较小,却显著抑制身不知的不定梢再生率,高浓度(≥300mg/L)的Carb不仅极显著抑制巴梨和身不知的出愈率,而且完全抑制了2者的不定梢再生;Km浓度大于10mg/L时,明显抑制愈伤组织的生长,且随着浓度的增加,死亡率逐渐增大,当Km浓度达100mg/L时,巴梨和身不知叶片死亡率达100%和96.30%。     

In vitro propagation of garlic by shoot proliferation

Shoot buds (5–8 mm long), excised from dormant cloves of the New Zealand commercial garlic (Allium sativum L.) and a virus-free French cultivar ‘Rose-de-Kakylis’, proliferated both axillary and adventitious shoots on B-5 basal medium supplemented with 0.5 mg l^?1 isopentenyladenine (2-ip) and 0.1 mg l^?1 naphthaleneacetic acid (NAA). An 8-fold increase in shoot number occurred every 6 weeks. Shoots were readily rooted in B-5 + 0.01 mg l^?1 2-ip + 0.2 mg l^?1 NAA and transferred to pots, where about 70% of the shoots formed established plants. The plants raised by this shoot-proliferation method retained the diploid condition of the parents.     

无籽西瓜子叶离体培养及植株再生试验

无籽西瓜比普通二倍体西瓜糖度高、食用方便、耐贮运，倍受人们青睐。但其制种复杂，成本高，所得的三倍体无籽西瓜种子中很多胚芽发育不良，子叶畸形，种皮厚，种子发芽率低，苗期生长缓慢，这些因素制约了无籽西瓜的生产。西瓜组织培养始于20世纪70年代，自1979年获得初步成功以来，国内外相继报道了这方面的研究进展，但利用无籽西瓜子叶作为外植体获得再生植株的报道甚少。为此，我们以无籽西瓜子叶为外植体，研究了通过愈伤组织阶段诱导芽及获得再生植株的培养条件。     

辣椒子叶和下胚轴的离体培养及高效再生体系的建立

采用三个辣椒品种的子叶和下胚轴,分别离体培养在附加不同激素及化合物的MB5培养基上,对苗龄、基因型、不同外植体和激素组合等对外植体不定芽诱导分化和芽伸长的影响进行研究。结果表明,苗龄对外植体不定芽分化的方式有直接影响;子叶的不定芽分化率高于下胚轴;B5维生素有利于芽的生长和芽伸长率提高。通过比较,筛选出了辣椒子叶和下胚轴离体再生的较好芽分化培养基为MB5＋6--BA5mg／L＋IAA0．5mg／L＋AgNO3 4mg／L＋蔗糖30g／L＋卡拉胶7g／L;芽伸长培养基为MB5＋6-BA3mg／L＋IAA1mg／L＋GA3 2mg／L＋蔗糖30g／L＋卡拉胶7g／L;生根培养基为1／zMS＋IAA0．2mg／L＋NAA0．1mg／L。     

In vitro somatic embryogenesis from Mangifera indica L. callus

The nucellus and globular adventitious proembryos were removed from 2-month-old fruits of mango (Mangifera indica L.) cultivars ‘Ono’ and ‘Chino’, and were cultured on sterile, solid Murashige and Skoog (MS) medium that had been modified as follows: half-strength major salts and chelated iron; 20% (v/v) coconut water (CW); 6% sucrose; 100 mg l^?1 ascorbic acid and 400 mg l^?1 glutamine. Embryogenic explants were sub-cultured after 4–6 weeks in liquid modified MS medium containing 2 mg l^?1 2,4-dichlorophenoxyacetic acid (2,4-D) instead of CW. Rapidly growing cultures were established and were sub-cultured monthly. Somatic embryogenesis was induced following sub-culture from MS medium with 2,4-D to MS without growth regulators and with or without activated charcoal (0.5%). Germination of somatic embryos appeared to be enhanced by 1 mg l^?1 benzyladenine (BA); however, most of the germinating embryos became embryogenic.     

In vitro regeneration from alginate-encapsulated microcuttings of Quercus sp.

Microcuttings with apical and nodal buds from common oak (Quercus robur L.) and Turkey oak (Quercus cerris L.) in vitro clones were encapsulated into an alginate matrix. Apical segments of common oak showed significantly greater in vitro regeneration potential as compared with the nodal ones. Encapsulated Turkey oak nodal segments demonstrated successful regeneration after different periods (2–6 weeks) of cold storage (4 °C), showing no significant deviation from the regenerative frequency of non-cold-treated segments.     

余甘子下胚轴离体培养中的器官形成与植株再生(英文)

以余甘子(phyllanthusemblicaL.)下胚轴为外植体,通过器官发生途径诱导形成不定芽,探讨不同激素组合对下胚轴器官发生和植株再生的影响,以期建立有效的再生体系,为以后的遗传转化研究奠定基础。结果表明,在附加0.1μmol/LNAA、5μmol/LBA和30g/L蔗糖MS培养基上培养,5周后不定芽再生频率达87.5%,平均每个外植体再生芽数为9.7个,为最佳组合。将分化的不定芽转移至含有10μmol/LIBA的MS培养基上,5周后生根,发育成健康的植株,生根率在29.2%。     

香榧茎段离体培养再生植株的研究

对香榧茎段不定芽诱导研究的结果表明,不定芽诱导的最佳培养条件为:培养基为B5+KT0.1mg/L+IBA0.5mg/L+0.08%活性炭+2%葡萄糖,培养温度为20℃,光强不高于400lx,该条件下的不定芽诱导率可达55%。将不定芽培养在1/2B5+IBA0.1mg/L+0.08%活性炭+2%葡萄糖的生根培养基上,生根率最高达到40%。通过器官发生途径国内外首次建立了香榧离体再生体系和快繁体系,并获得了完整的再生植株,为今后开展香榧的遗传转化和品种改良奠定了基础。     

阔叶猕猴桃叶片离体器官发生和植株再生(英文)

以阔叶猕猴桃(A ctinidia latifolia M err.)叶片为外植体,通过器官发生途径诱导形成不定芽,探讨了不同激素组合、暗培养时间以及不同浓度的蔗糖对叶片器官发生和植株再生的影响。结果表明,BW +0.1μm ol/L N AA +5μm ol/L Zeatin 附加20g/L 蔗糖,先进行21d 暗培养然后转到光下培养效果最好,6周后不定芽再生频率达91.67%,平均每个外植体再生芽数6.87个。再生芽生根良好,生根率可达100%。首次报道了阔叶猕猴桃的器官发生和植株再生,建立了叶片的高效再生体系,为今后的遗传转化研究奠定了基础。     

In vitro regeneration and Agrobacterium mediated transformation in gladiolus

Summary

In vitro regeneration and transformation studies were conducted on two cultivars of gladiolus. Cormels of 1.0 to 1.5 cm diameter cut into 2–3 mm thick slices of top, middle and bottom, and in vitro derived bisected shoot tips were used as explants on MS medium supplemented with 18.6 μM kinetin for multiple shoot induction. Amongst the cormel slices, the top slice gave better shoot induction response of 89% with an average of 2.4 shoots per explant over both cultivars. In vitro derived bisected shoot tips were inoculated on the medium oriented cut-side up, cut-side down and vertically both with and without the cormel base attached. Bisected shoot tips without attached cormel base and inoculated in the cut-side down orientation showed an average of 90% shooting response. In vitro derived shoot tips were used as explants for transformation. Explants were wounded by scalpel and particle bombardment with 1.6 μm naked gold particles by the biolistic delivery system. The wounded explants, after 3 d of recovery period, were co-cultivated with Agrobacterium strain LBA4404 harbouring the binary vectors pBI141 and pTOK233 which contained gus reporter gene with rice actin and 35S promoters respectively. GUS expression frequencies of 5.3% and 23% was obtained from scalpel and particle bombardment wounded explants, respectively. Particle wounded explants showed an average of 63 and 103 GUS spots when co-cultivated with pBI141 and pTOK233 binary vectors respectively. Explants co-cultivated with pBI141, after three weeks of selection on antibiotic containing medium showed blue streaks of GUS expression. It was concluded that Agrobacterium could infect the monocot gladiolus and transform the tissue eficiently when tissues were prewounded with naked gold particles delivered by particle gun.     

苦瓜下胚轴的离体再生体系

为了建立高频的苦瓜离体再生体系,为苦瓜遗传转化提供技术基础,筛选了最适消毒方法和外植体类型。以MS培养基为基础培养基,添加不同浓度激素组合,筛选最适于苦瓜不定芽诱导、伸长和生根的培养基。最佳的消毒方法为:用水浸泡去壳的种子15 min,在超净工作台中用75%酒精浸泡30 s后再用4%NaClO浸泡6 min,用无菌水冲洗4～5遍,污染率为0,发芽率为98%;最佳的愈伤组织诱导培养基为MS+2.5 mg·L~(-1)6-BA+0.01 mg·L~(-1)IBA,愈伤组织密度为0.79 g·cm~(-3),不定芽的分化率为39%,最佳外植体为下胚轴(12 d),诱导率为41%;最佳的增殖培养基为MS+1.0 mg·L~(-1)6-BA+0.005 mg·L~(-1)IBA;最佳的生根培养基为MS+0.1 mg·L~(-1)IBA,移栽存活率为74%。初步建立了苦瓜离体再生体系。     

寒富苹果叶片离体再生及四倍体诱导

为建立寒富苹果高效的离体再生体系和多倍体诱导体系,以试管苗叶片为外植体,研究了培养基中激素对寒富叶片再生芽的影响及适宜的四倍体诱导方法。结果表明,当培养基中BA质量浓度为0.5mg/L时,再生频率最高达35.7%;当培养基中BA质量浓度为2.5mg/L时,再生频率超过90%,平均再生芽数在4以上。在培养基中附加1.0mg/LTDZ,再生频率达100%,平均再生芽数达19.47。以附加15、30、60、120mg/L秋水仙素的液体再生培养基处理叶片5d,各个质量浓度处理均诱导出四倍体植株,诱变率在5.3%～22.2%之间;寒富苹果叶片在附加50mg/L秋水仙素固体再生培养基上处理5d亦获得了四倍体植株。研究结果表明寒富苹果叶片具有极强的不定芽再生能力,叶片再生过程中进行秋水仙素处理是获得其四倍体的一个有效途径。     

美人指葡萄不定芽离体诱导再生植株的研究

6-BA、TDZ与IBA不同浓度组合对美人指葡萄叶片、叶柄和茎段不定芽离体器官发生途径诱导再生植株有不同的影响。以茎段为外植体在MS+6-BA1.0mg/L+IBA0.01mg/L培养基上分化效果最好,再生率75.00%;叶片、叶柄分别在MS+6-BA3.0mg/L+IBA0.10mg/L和MS+6-BA2.0mg/L+IBA0.05mg/L培养基上分化效果较好。TDZ和IBA研究的几种组合都能诱导不定芽再生,但再生率不高。     

影响甘蓝型油菜下胚轴外植体芽高频率再生的因素

以优质高产杂交油菜新品种"蜀杂九号"父本84100-18(BrassicanapusL)油菜品系的下胚轴为外植体,分析讨论了预培养,苗龄,激素浓度,AgNO3等因素对下胚轴的再生条件的影响.试验表明:7 d～9 d(天)的苗龄的外植体出愈率很高;MS 3 mg/L(毫克/升)6-BA时芽的分化率最高;AgNO3可提高下胚轴芽的分化率;提高琼脂浓度,降低蔗糖含量可以减少玻璃化现象.     

黑莓试管苗叶片植株的再生

以黑莓带芽茎段试管苗叶片为外植体,诱导形成不定芽并进一步形成再生植株。在MS+TDZ2.0mg/L的培养基中叶片不定芽最高分化频率达93.7%(6.52不定芽/外植体),在不定芽伸长培养基BA0.5mg/L上,分化完全的不定芽能够长大伸长,当其高度达到4cm时,切下转接于IBA为0.3mg/L培养基上诱导生根,生根率100%。     

丽江山定子15个矮生单系的离体培养

丽江山定子 ( Malusrockii Rehder)原产我国云南及四川和西藏 ,喜湿耐涝 ,与苹果嫁接亲和力强。 1987年以来 ,我们采用过氧化物酶同工酶检测与形态特征鉴定评价相结合的方法 [1 ] ,从 8463 6株原产于云南省的丽江山定子1年生实生苗中 ,选出了 15个矮生单系。经过8年的观察和鉴评 ,发现其中 9个矮生单系母株 1年生枝条的长度仅为 0 .9～ 6.4cm,且 1年生枝条和 2年生枝条的总长度也只有 1.9～10 .2 cm。经扦插试验 ,这 15个矮生单系枝条扦插极难生根 ,很难进一步繁殖用作矮化中间砧的试验试材。经过离体培养试验 ,我们筛选出了适合于这 15个…     

In vitro regeneration and conservation of wild species of Arachis

Summary

Wild species of Arachis are restricted to South America and generally occur in regions under intensive environmental disturbance. Both in situ and ex situ conservation strategies are required in order to maintain the availability of these genotypes. This work developed in vitro regeneration systems from seed explants of 17 wild species of Arachis from six Sections (Heteranthae, Caulorrhizae, Triseminatae, Erectoides, Procumbentes and Arachis). After seed disinfection, embryonic axes, leaflets and cotyledons were excised aseptically and cultured on Murashige and Skoog (MS) medium supplemented with 8.8 µM, 22 µM or 110 µM 6-benzylaminopurine (BAP). Cultures were maintained in a growth chamber at 28° ± 2°C with a 16 h photoperiod. Regeneration patterns from seed explants were similar among species from all Sections. Embryonic axes produced plants through meristematic amplification or multiple shoot formation, while cotyledons and embryonic leaflets produced shoots at significantly lower frequencies through direct and indirect organogenesis, respectively. Shoots obtained from all explants were transferred to MS medium without growth regulators to induce root formation.     

Plantlet regeneration from root callus of different Prunus species

Caulogenesis was obtained in vitro on the calli of Prunus sp., P. dawyckensis, P. canescens and hybrid P. incisa x serrula. These calli were formed on the roots of plantlets derived from meristem culture on a medium containing 6-benzylaminopurine (BAP) (10^?3 g/l) and gibberellic acid (GA₃) (10^?4 g/l). The first buds appeared after a month. After transfer of the calli on the same medium, the caulogenesis continued. The degree of regenation varied according to the species.     

秋福红树莓叶片离体再生植株研究

以红树莓品种秋福(Rubus L.cv.Autumn Bliss)离体叶片为外植体,研究适合其再生植株的叶片部位、放置方式、植物生长调节剂以及暗培养时间等条件。结果表明,叶片外植体接种于MS+TDZ2.00mg/L+IAA0.10mg/L的培养基,暗培养2～3周转移至正常光照下培养效果最好,愈伤组织形成率、不定芽再生率和外植体不定芽数分别为100.00%、95.83%和(5.57±0.27)个。将再生芽接种于1/2MS+IBA0.10mg/L的培养基中,35d后生根率达100.00%。再生植株炼苗后移栽,30d时成活率达97.30%,成功地建立了红树莓叶片的离体再生体系。     

Growth and regeneration of plantlets in callus cultures of pineapple

Callus cultures were established from the basal region of in vitro-obtained shoot buds on MS medium supplemented with CH + CW + NAA. Such callus cultures, when grown on MS medium devoid of any growth regulators, regenerated shoot buds and optimum regeneration was obtained on MS + CW (5% v/v) medium. Addition of BA did not enhance shoot bud regeneration, but two variants (albino types) were observed among the BA-induced regenerants. The callus-regenerated shoot buds produced multiple shoots when transferred to MS + NAA + IBA + K medium. The plantlets were induced to root on a modified White's medium + NAA + IBA and subsequently transferred to soil.