Somatic embryogenesis and histological analysis from zygotic embryos in Vitis vinifera L. ＇Moldova＇

We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera L. 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) medium supplemented with 1.0mg'L-1 2,4-D and 0.5 mg'L-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg-L-1 6-BA and 2 mg.L-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg.L-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dy-namic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic em-bryos, including typical epiderma, cotyledon primordium and vascular tissue.     

Somatic embryogenesis and histological analysis from zygotic embryos in Vitis vinifera L. ＇Moldova＇

We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape ＇Moldova＇ （Vitis vinifera U ＇Moldova＇）. Primary calli were initiated on Nitsch and Nitsch （NN） medium supplemented with 1.0 mg·L^-1 2,4-D and 0.5 mg·L^-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg·L^-1 6-BA and 2 mg·L^-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg·L^-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dynamic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic embryos, including typical epiderma, cotyledon primordium and vascular tissue.     

花曲柳体胚发生和植株再生

以花曲柳合子胚的单片子叶为外植体成功诱导出体胚并获得再生植株。未成熟合子胚的子叶在添加400mg·L-1水解酪蛋白、0.25mg·L-16-BA、1.5mg·L-1NAA、70g.L-1蔗糖和6g·L-1琼脂的MS1/2培养基上可以成功诱导产生体胚,诱导率达到34.7%,每个外植体上体胚数量为2～9个。成熟合子胚的子叶在添加0.25mg·L-16-BA、2mg·L-1NAA的MS1/2培养基(其他成分同上)上可以成功诱导产生体胚,诱导率为10.0%。体胚在MS1/2培养基上经过成熟培养后可以正常萌发,萌发率87.6%。萌发的体胚植株在MS1/2+0.01mg·L-1NAA培养基上生长较好,具备实生幼苗的外观特征。经炼苗后的体胚苗移植到栽培基质(草炭土:蛭石:珍珠岩体积比为5:4:1)中可以正常生长,成活率为75.0%。     

刺五加体细胞胚状体发生及成苗的研究

以刺五加试管苗叶片为试材,将叶片接种于含有不同种类和浓度激素的培养基上进行培养,研究不同激素对刺五加胚性愈伤组织诱导、胚状体发生及生根的影响。结果表明：诱导叶片胚性愈伤组织发生的最适培养基为MS＋6-BA 1.5 mg.L-1＋2,4-D 1.0 mg.L-1,胚状体生长发育的最适培养基为MS＋6-BA1.0 mg.L-1＋IBA0.5 mg.L-1,生根的最适培养基为1/2MS＋NAA0.4 mg.L-1。     

Somatic embryogenesis and plant regeneration inAcanthopanax sciadophylloides

Somatic embryos ofAcanthopanax sciadophylloides Franch. et Sav. were differentiated from both zygotic and somatic embryos and calli, and plants were regenerated from these somatic embryos. A zygotic embryo, enclosed within a small portion of the endosperm, was incubated on Murashige and Skoog (MS) media supplemented with various combinations (range 0–10.0 mg/l) of 6-benzylaminopurine (BAP) and 2,4-dichlorophenoxyacetic acid (2,4-D). After 4 months, swelling of the zygotic embryos and callus formation was observed. When the swollen embryos were transferred to MS medium supplemented with 0.5 mg/l of 2,4-D, somatic embryos were formed in one to two months. After subculture on the same medium, new embryos were differentiated from various parts of the older somatic embryos. The calli were cultured on medium supplemented with 2.0 mg/l of 2,4-D and BAP for three weeks. Proliferated calli were transferred to medium supplemented with 1.0 mg/l of 2,4-D and BAP. Somatic embryos were differentiated from the calli within one to two months. Somatic embryos were germinated on half-strength MS medium without plant growth regulators and the plantlets were grown in soil. A part of this paper was presented at the 106th Annual Meeting of the Japanese Forestry Society (1995) & First Asia-Pacific Symposium on Forest Tree Genetic Improvement (Beijing).     

刺槐未成熟合子胚的体细胞胚胎发生和植株再生

以刺槐不同胚龄的未成熟合子胚为外植体,采用混合正交试验设计,研究幼胚胚龄和不同外源激素种类及质量浓度对刺槐胚性愈伤组织的诱导和体细胞胚分化、萌发的影响.结果表明:开花后8周(55天左右)是胚性愈伤组织和体胚诱导的最佳外植体取材时期;MS+2,4-D 5.0 mg·L-1 +BA0.5 mg·L-1是诱导胚性愈伤组织的最佳培养基,出愈率最高为95.42％±0.02％;MS +NAA0.5 mg·L-1 +BA0.5 mg·L-1+谷氨酰胺250 mg·L-1+水解酪蛋白500 mg·L-1是体细胞胚诱导和分化的最佳培养基,直接体细胞胚发生率最高为92.40％±0.12％,通过愈伤组织诱导体细胞胚发生的频率最高为90.73％ ±0.49％.一旦形成球形胚,将培养物转接到不含任何生长调节剂的MS培养基上,体细胞胚经成熟萌发可进一步形成完整小植株.     

红松愈伤组织诱导的初步研究

以未成熟和成熟合子胚为外植体进行了红松愈伤组织的诱导,其结果表明,成熟胚的诱导效果明显好于幼胚;鉴于诱导产生褐化的主要部位来自胚根,所以外植体应选用子叶、胚轴或去除胚根的胚;诱导培养基采用LM＋2,4-D2．0mg／L＋6-BA0．5mg／L可诱导出较大量Ⅱ型有效愈伤组织,经组织学观察,可见早期原胚团,疑似胚性愈伤组织;继代增殖培养基可采用LM＋2．4-D0．5mg／L＋6-BA0．2mg,／L。     

香蕉薄切片愈伤组织再生体系的建立研究

以巴西香蕉品种的薄切片为外植体高频地诱导出愈伤组织,并诱导出了不定芽,获得了再生植株。在实验过程中,利用正交实验中筛选出愈伤组织诱导的最佳培养基为MS+2,4-D 4.0 mg.L-1+KT 1.0 mg.L-1+NAA 1.0 mg.L-1+活性炭200 mg.L-1,诱导率可达100%,愈伤组织增殖的最佳培养基是MS+2,4-D 2.0 mg.L-1+6-BA 0.5 mg.L-1+NAA 0.5 mg.L-1+活性炭100 mg.L-1,继代3-4次的愈伤组织在6-BA浓度为5.0 mg.L-1的分化培养基上能分化出不定芽,继代10次之后愈伤组织在继代培养基上开始出现褐化现象,愈伤组织增殖明显受到抑制,褐化的愈伤组织逐渐死亡。     

离体培养条件下核桃器官发生和体细胞胚胎发生

采自幼树、成年树及成年树伐根萌条的嫩梢腋芽,在改良DKW培养基+6-BA1.0mg/L中可伸长生长;在加有6-BA1.0mg/L+IBA 0.01mg/L的继代培养基中,可保持腋芽的生长并形成不定芽。芽增殖率为每月600％左右。芽苗经5.0mg/L IBA处理7天,然后在含活性炭的无激素培养基中培养20天,有54％可生根成苗。5月中旬至6月上旬的幼胚,在改良DKW培养基+6-BA1.0 mg/L中,黑暗培养30天左右可产生体细胞胚,并可连续多代保持胚性分生能力。体胚在无激素的发芽培养基中黑暗培养7天左右可发芽成苗。胚性愈伤组织在悬浮振荡培养中可保持分生能力。     

利用悬浮培养和培养基选择改善火炬松的体细胞胚胎发生和植株再生(英文)

三个基固型的火炬松成熟合子胚被培养在附加 8mg·L-12 ,4 D ,4mg·L-1BA ,4mg·L-1KT ,5 0 0mg·L-1水解酪蛋白和 5 0 0mg·L-1谷氨酰胺的愈伤组织诱导培养基上诱导愈伤组织 .来自于子叶、胚轴和胚根的愈伤组织在附加 1 6mg·L-12 ,4 D ,0 8mg·L-1BA和 0 8mg·L-1KT的愈伤组织增殖培养基上培养 9周后 ,可获得 16 9%的胚性愈伤组织 .通过建立胚性细胞悬浮系和研究ABA、PEG和活性炭对体细胞胚成熟的促进作用 ,优化的体细胞胚胎发生体系被建立 .71棵再生小苗被用于移栽试验 ,2 3棵小苗在田间移栽成活     

印度萝芙木组织培养技术研究

以印度萝芙木枝条顶芽为外植体进行离体培养,外植体最佳灭菌方法为0.08%HgCl2处理时间20min;诱导愈伤组织培养基为MS+L-半胱氨酸0.02mg·L-1+6-BA2.0mg.L-1+NAA0.2mg·L-1,诱导率可达100%;诱导不定芽培养基为MS+L-半胱氨酸0.01mg·L-1+6-BA1.0mg·L-1+NAA0.1mg·L-1;生根培养基为1/2MS+NAA1.0mg·L-1,生根率可达100%。     

齿瓣石斛组织培养技术研究

以齿瓣石斛蒴果为外植体进行胚培养的试验表明,初始萌发培养基为1/2MS;原球茎萌发培养基为3/4MS+10%马铃薯汁+5%香蕉汁+6-BA0.2mg·L-1+NAA0.05mg·L-1+0.05%AC;继代增殖培养基为MS+10%马铃薯汁+10%香蕉汁+6-BA0.5mg.L-1+NAA0.2mg.L-1+0.1%AC;壮苗生根培养基为1/2MS+10%香蕉汁+5%马铃薯汁+6-BA0.2mg·L-1+NAA1.0mg·L-1+0.1%AC,生根诱导率达100%,成活率达90%以上。     

白刺花胚性愈伤组织诱导及体细胞胚发生

【目的】探讨不同植物生长调节剂对白刺花胚性愈伤组织诱导的作用,以及培养基中氮源和无机盐浓度对白刺花体细胞胚发生和植株再生的影响,以期建立白刺花体细胞胚发生、发育及调控技术体系,为白刺花种苗快速繁殖体系建立及遗传转化研究提供参考。【方法】以白刺花叶片为外植体,研究生长调节剂2,4-D(1.0、2.0、3.0、4.0 mg ·L -1 )、NAA(0、0.5、0.8、1.0 mg ·L -1 )、6-BA(0.2、0.5、1.0、2.0 mg ·L -1 )和TDZ(0、0.2、0.5、1.0 mg ·L -1 )组合对胚性愈伤组织诱导,及NAA(0、0.2、0.5 mg ·L -1 )、6-BA(0、0.5、1.0 mg ·L -1 )和TDZ(0、0.2、0.5 mg ·L -1 )组合对体细胞胚发生的调控作用,筛选最优生长调节剂组合;并研究培养基中KNO 3和NH 4NO 3比例对体细胞胚发生的作用,及MS培养基中无机盐浓度(1/5MS 、1/4MS、1/3MS、1/2MS)对体细胞胚萌发的影响,筛选最佳的体细胞胚发育及成熟萌发条件。【结果】白刺花叶片外植体胚性愈伤组织诱导适宜培养基为MS + 2,4-D 3.0 mg ·L -1 + NAA 0.5 mg ·L -1 + 6-BA 0.2 mg ·L -1 + TDZ 1.0 mg ·L -1 +蔗糖40 g ·L -1 +琼脂7.0 g ·L -1 ,诱导率为42.0%。采用MS基本培养基时,最佳的体细胞胚发生培养基为MS + NAA 0.5 mg ·L -1 + 6-BA 1.0 mg ·L -1 + TDZ 0.5 mg ·L -1 +蔗糖40 g ·L -1 +谷氨酰胺100 mg ·L -1 +琼脂7.0 g ·L -1 ,体细胞胚发生率为78.46%,总胚数为对照的3.6倍;MS培养基中,KNO 3浓度提高1倍、NH 4NO 3降至1/2时,体细胞胚发生率可提高至91.33%,总胚数为采用MS基本培养基时的1.4倍;1/3MS培养基有利于体细胞胚的萌发,萌发率为82.75%,幼苗长势良好,单株平均鲜质量为76 mg,幼苗驯化移栽1个月后成活率达90%以上。【结论】白刺花叶片接种于添加2,4-D、NAA、6-BA和TDZ不同组合的诱导培养基上,可脱分化形成愈伤组织或胚性愈伤组织,2,4-D浓度对愈伤组织形态和质地有较大影响。TDZ有利于体细胞胚的形成,适宜浓度的生长素与细胞分裂素组合及硝态氮和铵态氮的比例对体细胞胚的形成和发育具有调控作用,降低MS无机盐浓度可提高体细胞胚萌发率,本试验体系的再生植株移栽成活率达90%以上。     

栓皮栎体胚的增殖、成熟和萌发

以栓皮栎实生苗叶片为外植体诱导体细胞胚胎发生,调查碳水化合物、渗透剂和植物生长调节剂对体胚增殖、成熟和萌发的影响,建立体胚增殖、成熟和萌发的培养基方案.叶片外植体在附加1.0 mg·L-1NAA和0.5mg·L-1BA的起始培养基上诱导形成前胚性团块.这些胚性团块在增殖培养基上培养6周,在附加1 mg·L-1BA、0.25 mg·L-1NAA和3%蔗糖的MS培养基上增殖效果最好.将单个体胚转接到成熟培养基上进行培养,蔗糖浓度对栓皮栎体胚成熟以及后续的萌发有显著影响.成熟培养基中附加5%的蔗糖,体胚成熟率和萌发率分别达到63.5%和33.8%.虽然在成熟培养基中附加ABA有利于体胚成熟,但对体胚的进一步萌发没有促进作用.为了提高萌发率,成熟体胚在附加植物生长调节剂的萌发培养基上进行培养,以及进行预冷处理.成熟体胚4℃冷处理没有促进胚根和上胚轴的发育萌发.在附加0.5 mg·L-1BA和0.25 mg·L-1 IBA的1/2 MS萌发培养基上,体胚萌发率达到65.9%,再生植株转化率达到9.4%.     

Regeneration of <Emphasis Type="Italic">Melia volkensii</Emphasis> Gürke (Meliaceae) through direct somatic embryogenesis

Experiments were conducted to study plant regeneration through direct somatic embryogenesis using mature zygotic embryo and cotyledonary explants from seeds of Melia volkensii stored for <3 and >12 months. Explants were cultured on Murashige and Skoog (MS) medium supplemented with BAP, NAA and 2,4-D (0.5, 1.0 and 2.0 mg l⁻¹) alone, and BAP (0.5, 1.0, 2.0 and 4.0 mg l⁻¹) in combination with 2,4-D or NAA (0.2 and 0.5 mg l⁻¹). After 4 weeks in culture, up to 60% of cotyledonary explants from the seeds stored for <3 months produced direct somatic embryos on BAP (0.5–4.0 mg l⁻¹) in combination with 2,4-D (0.2 mg l⁻¹). The number of somatic embryos ranged from 5 to 14 per explant in BAP (0.5 mg l⁻¹) and 2,4-D (0.2 mg l⁻¹) combination. Only 20% of cotyledonary explants from seeds stored for >12 months produced somatic embryos. Mature zygotic embryos failed to produce any somatic embryos. Subcultures of somatic embryos from cotyledonary explants of seeds stored for <3 months formed clusters of shootlets on semi solid MS and 1/2 MS media. After 6 weeks of subculture on multiplication MS media augmented with BAP (0.5 mg l⁻¹) and IAA (0.2 mg l⁻¹), 70% of the shoot tips formed 4–7 shoots per explant. Up to 33% of the multiplied shoots were rooted in MS medium supplemented with 2.0 mg l⁻¹ IBA. Plantlets developed normally into seedlings in the greenhouse.     

红松成熟合子胚愈伤组织诱导的研究

以红松(Pinus koraiensis Sieb.et Zucc.)成熟合子胚为基准试验材料,对红松愈伤组织诱导的影响因素进行了初步研究。结果表明,不同的培养基对有效愈伤组织的诱导效果不同,初始诱导应选用LM培养基使外植体完全脱分化,反复继代后可以产生有效愈伤组织,供试的其他培养基继代后则极少产生有效愈伤组织;不同的激素组合和浓度配比对有效愈伤组织的诱导率影响不同,2,4-D是愈伤组织启动的不可或缺的植物生长调节剂,其作用远远大于NAA。在LM+2mg.L-1 2,4-D+0.5mg.L-16-BA的诱导培养基中加入0.5mg.L-1 KT或0.5mg.L-1 TDZ,可明显提高有效愈伤组织的诱导率(可达75.7%)。     

两种优良巴西杂交桉树的组织培养和快速繁殖

本研究以巴西尾巨桉和巨赤桉两种优良杂交桉树的带芽茎段为外植体进行离体培养和快速繁殖研究.结果表明:尾巨桉和巨赤桉再生芽诱导的最佳培养基为改良MS+6-BA 0.5 mg·L-1+NAA 0.2 mg·L-1,继代增殖的最适培养基为改良MS+6-BA 0.5 mg·L-1+NAA 0.15 mg·L-1.芽诱导和继代增殖中6-BA的浓度对两种杂交桉树有显著影响.生根培养中,尾巨桉和巨赤桉生根情况差异显著,尾巨桉茎芽最适生根培养基为1/2MS+NAA0.2 mg·L-1+IBA 1.0 mg·L-1,巨赤桉茎芽最适生根培养基为1/2MS+NAA 0.15 mg·L-1+IBA 1.0 mg·L-1.两种杂交桉树组织培养体系的建立,为桉树良种选育和遗传转化体系的研究提供技术参考.     

大岩桐组培育苗试验

以大岩桐幼嫩叶片、老叶、带腋芽的茎段为外植体,进行试管诱导,并进行继代、生根、移栽试验.结果表明:大岩桐诱导外植体以带腋芽的茎段效果最好;适宜的启动培养基、增殖培养基和生根培养基分别为MS+6-BA 1.0mg· L-1+NAA0.2mg·L-1、MS+6-BA0.5 mg·L-1 +NAA0.1 mg·L-1和1/2...     

钟花樱组织培养繁殖研究

以钟花樱(Cerasus campanulata)的嫩枝作外植体进行组织培养技术研究。结果表明：适合不定芽诱导的培养基为 MS+6-BA 1.0 mg · L-1+NAA 0.1 mg · L-1+蔗糖30 g · L-1+卡拉胶6.5 g · L-1,诱导率为44.44%,适合增殖培养的培养基为 MS+6-BA 1.0 mg · L-1+NAA 0.05 mg · L-1+GA30.5 mg · L-1+蔗糖30 g · L-1+卡拉胶6.5 g · L-1,增殖倍数为3.70,适合生根培养的培养基为1/2 MS+IBA 1.5 mg · L-1+NAA 0.1 mg · L-1+蔗糖30 g · L-1+卡拉胶6.5 g · L-1+活性碳0.5 g · L-1,生根率为82.23%。