诱导不同基因型大豆未成熟子叶体细胞胚胎发生的研究

以未成熟子叶为外植体,研究25种基因型对大豆愈伤组织形成与体细胞胚胎发生的影响。结果表明,大豆不同基因型间愈伤组织诱导率变化较小,而基因型对体细胞胚胎发生的影响较大,依基因型不同体细胞胚诱导率变化在3.33%～83.33%。体细胞胚诱导率、平均胚数在基因型间的差异均达极显著水平,且两者间存在极显著相关关系。     

毛白杨PtAUX1超表达诱导体状胚发生

体状胚的发生由一个生长素信号级联系统控制,其发生过程需要大量的生长素,随后在去除生长素或降低生长素浓度的条件下,体状胚趋于成熟。除植物激素外,环境胁迫也是体状胚发生所必须的,激素和环境胁迫协同诱导细胞的脱分化、再分化和体状胚的发生。目前发现的参与体状胚发生的主要基因有SERK、PKL和BBM等,它们作用于体状胚的不同发育阶段。本研究中,在毛白杨35S::PtAUX1超表达植株中发现叶片表面产生大量体状胚,体状胚分散在叶片主脉及侧脉两侧或聚集在叶片边缘,其中部分体状胚可发育成成熟体状胚。将成熟体状胚取下,置于MS0培养基上进行培养,体状胚发芽并缓慢产生根。研究结果表明,PtAUX1通过建立新的生长素浓度梯度,诱导体细胞转化成胚性细胞,从而使叶片产生出体状胚。除上述基因外,PtAUX1也参与体状胚的发生,这一研究发现现尚未见报道。     

Effect of incubation temperature regimes and culture medium on broccoli microspore culture embryogenesis

Conditions for reliable induction of embryogenesis from isolated microspores were studied in ten genotypes of broccoli. Embryo yields were significantly increased in almost all of the broccoli genotypes by the incubation at 32.5 ^°C for 1 day, than when the standard incubation at 30 ^°C for 2 days was used. Treatments of 48 hours at 32.5 ^°C produced less than optimal results suggesting that broccoli microspores are more sensitive to high temperatures than those of B. napus. The use of the ¹/₂ NLN-13 medium yielded greater number of embryos than the standard NLN-13. The magnitude of the response to the redution of the concentration of major salts by half in the NLN medium varied with the different genotypes. High embryogenic broccoli cultivars, such as ‘Shogun’, ‘SDB9’, and ‘Green Valiant’, presented a better response to the reduction of the concentration of major salts by half in NLN-13. Reduction never produced a detrimental effect on embryo yield and seems not to have any effect in the subsequent development of embryos in plants. This revised version was published online in July 2006 with corrections to the Cover Date.     

体细胞克隆牛0～12月龄生长的测定研究

通过对5头体细胞克隆母犊牛及其同期自然繁殖出生的荷斯坦母犊牛初生期、6月龄、12月龄的体重、体尺指标测定,初步分析了其生长发育速度的变化规律。结果表明:(1)克隆牛初生重53.67±7.00kg,明显高于自繁奶牛(P<0.01),6月龄体重141.00±16.49kg,明显低于自繁奶牛(P<0.01),12月龄体重302.80±32.17kg,与自繁奶牛基本相当;(2)克隆牛0～6月龄期间平均日增重0.70±0.11kg明显低于自繁奶牛(P<0.01),6～12月龄期间平均日增重0.90±0.12kg,与自繁奶牛基本相当(P>0.05),0～12月龄期间平均日增重0.70±0.11kg,与自繁奶牛基本相当(P>0.05);(3)克隆牛初生体尺指标明显大于自繁奶牛(P<0.01),6月龄体尺指标明显小于自繁奶牛(P<0.01),12月龄体尺指标与自繁奶牛相当(P>0.05);(4)克隆牛0～6月龄期间体尺平均增长明显低于自繁奶牛(P<0.01),6～12月龄期间体尺平均增长明显高于自繁奶牛(P<0.01),0～12月龄期间体尺平均增长明显低于自繁奶牛(P<0.01)。     

植物生长调节剂在植物体细胞胚发生中的应用

合理应用植物生长调节剂是成功诱导体细胞胚的决定因素,也是影响体细胞胚高频率发生的主要因素。本文从生长素类、细胞分裂素类、赤霉素、脱落酸以及各类植物生长调节剂间组合使用等几个方面综述了外源植物生长调节剂在体细胞胚发生中的应用进展,为植物体细胞胚的诱导提供了一定的参考价值。     

The Prospects of Tissue Culture and Genetic Engineering for Strawberry Improvement

Abstract

Strawberry shoot meristem cultures have been used for producing virus-free strawberry plants. Plantlet regneration from leaf mesophyll protoplasts, and Agrobacterium tumefaciens mediated genetic transformation, mark the beginning of strawberry improvement with biotechnologies. The use of biotechnological tools has a greater potential in producing strawberry plants with traits such as early maturation, frost and disease resistance, mite and nematode resistance, high yield, better quality, suitability for mechanical harvesting, shipping ability, ellgaic acid content, aroma compounds and cost-effective micropropagation. Gene identification and mapping with RFLPs or RAPDs would be desirable. Transgenic strawberries should be tested for their quality before releasing to the consumers.     

枣幼胚体细胞胚的发生及植株再生

以小梨枣胚龄30~40d的幼胚为外植体,研究不同质量浓度6-苄基腺嘌呤(6-BA)、噻苯隆(TDZ)、吲哚丁酸(IBA)、萘乙酸(NAA)、脱落酸(ABA)及其组合对幼胚胚乳看护培养效果、体细胞胚发生、成熟及萌发的影响。结果表明,适宜小梨枣幼胚(30~40d)胚乳看护培养的培养基为MS+IBA0.2mg·L~(-1)+BA0.5mg·L~(-1)+NAA 0.1 mg·L~(-1),成胚率达62.07%;诱导胚性愈伤组织较适宜的培养基为MS+TDZ 0.6mg·L~(-1)+IBA1.5mg·L~(-1)+NAA 0.1mg·L~(-1),诱导率为53.32%;在MS+6-BA 0.3mg·L~(-1)+NAA0.02mg·L~(-1)组合下有利于体细胞胚的发生,发生率可达71.43%;ABA可促进体胚的正常发育,质量浓度为0.5mg·L~(-1)时,体细胞胚结构完整,体胚成熟率为21.83%;将成熟的体胚转接到含有IBA 0.03mg·L~(-1)+GA30.03mg·L~(-1)的培养基中,体细胞胚萌发形成完整植株。     

水曲柳体胚发生过程中不同状态类型外植体的生理生化状态

以水曲柳Fraxinus mandshurica幼嫩合子胚子叶为外植体诱导水曲柳体胚发生,对该过程中外植体的不同状态类型分别取样,检测这些不同状态类型外植体的多糖含量、可溶性蛋白质含量及超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、多酚氧化酶(PPO)活性等生理生化指标,分析不同状态类型外植体生理生化与体胚发生、愈伤组织发生、外植体褐化、外植体生长等方面的关系,以便为分析水曲柳体胚发生机理和改善其体胚发生技术提供依据.研究结果表明,不同状态类型外植体的生理生化状态与体胚发生、愈伤组织发生、外植体褐化和生长发育存在非常有趣的相关关系,值得进一步深入解析.     

原生质体电融合再生柑橘属间四倍体体细胞杂种

自 Ohgawara等 (1 985)得到枳与 Trovita甜橙属间体细胞杂种植株近 1 5年来 ,已得到了约 1 50例柑橘体细胞杂种〔3〕,其中有种、属间有性和嫁接亲和的组合 ,也有有性和嫁接均不亲和的属间、族间组合。这些体细胞杂种的获得 ,丰富了柑橘的种质资源库 ,为育种提供了可供选择和利用的新型试材。本文报道的是最近获得的一例金柑与柑橘属间体细胞杂种。1　材料与方法1 .1　材料丹西红橘 (Citrusreticulata )愈伤组织悬浮系继代 3次后 ,于第 7～ 1 0天时取愈伤组织分离原生质体 ;长寿金柑 (Fortunella obovata )种子播种于 MT基本培养基 (含 3 …     

Somatic embryogenesis and Agrobacterium-mediated transformation of Japanese apricot (Prunus mume) using immature cotyledons

This study describes a successful method of somatic embryogenesis and genetic transformation using immature cotyledons of Prunus mume. Immature cotyledons from four different developmental stages of eight different P. mume cultivars were used for the experiments to optimize somatic embryogenesis and genetic transformation protocols. Somatic embryogenesis was induced when the explants were cultured on somatic embryo inducing medium consisting of MS basic medium supplemented with 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 μM 6-benzyladenine (BA). They were cultured for 30 days and then transferred to somatic embryo propagation medium containing 0.1 μM α-naphthaleneacetic acid (NAA) and 5 μM BA. It appeared that the developmental stage of the immature cotyledons used as explants was the most important factor for somatic embryogenesis; higher frequencies of somatic embryogenesis were observed when the immature cotyledons were less than 5 mm in length regardless of cultivars. For genetic transformation, the immature cotyledons were inoculated with Agrobacterium tumefaciens EHA101 harbouring a binary plasmid vector with neomycin phosphotransferase II and an intron-interrupted β-glucuronidase gene under the control of cauliflower mosaic virus 35S promoter, and three transgenic plant lines were obtained from inoculated “Sirakaga” immature cotyledons. Transgenic somatic embryos and shoots were selected using 25 mg l⁻¹ kanamycin. Integration of transgenes in the genome of GUS-positive putative transgenic shoots was confirmed by PCR and Southern blot analyses.