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青扦胚性细胞悬浮培养中影响体细胞胚发生因素的研究 总被引:4,自引:0,他引:4
试验以青扦(Piceawilsoni)的胚性愈伤组织为材料,以改良59基本成分附加24-D1mg/L及KT1mg/L为培养介质,比较了液体悬浮与半固体二种培养方式对胚性愈伤组织增殖和体细胞发生的影响,研究了液体悬浮培养过程中影响体细胞胚发生的因素。结果表明:液体悬浮培养好于半固体培养,它的胚性愈伤组织的生长率为268%,是半固体培养的124倍;体细胞胚的分化率为93%,是半固体培养的22倍;悬浮培养较佳的培养条件为:初始细胞密度为2%(鲜重),蔗糖浓度为20g/L,摇床转速为100r/min,pH为58。经过两个月悬浮培养,将培养物转至1/2改良59附加ABA1mg/L的分化培养基上,3个月后每g培养物上可获得285个正常的子叶期体细胞胚。 相似文献
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对12个品种的二棱大麦幼穗和6个品种的幼胚在附加500mg/L酪蛋白,150mg/L天门冬素和2mg/L 2,4-D的MS培养基(M320)中进行了离体培养,并诱导产生出愈伤组织。把由D1幼胚所获得的愈伤组织转移到含2,4-D0.5mg/L(M305)培养基中后诱导了产生出了体细胞胚性愈伤组织,幼苗和根。筛选出了体细胞胚性无性系。在长达二年的继代培养中,该细胞系仍保持旺盛的再生和分化成苗能力。 相似文献
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对12个品种的二棱大麦(Hordeumdistichum2n=24)幼穗和6个品种的幼胚在附加500mg/L酪蛋白,150mg/L天门冬素和2mg/L2,4-D的MS培养基(M_(320))中进行了离体培养,并诱导产生出了愈伤组织。把由D_1幼胚所获得的愈伤组织转移到含2,4-D0.5mg/L(M_(305))培养基中后诱导产生出了体细胞胚性愈伤组织,幼苗和根。筛选出了体细胞胚性无性系。在长达二年的继代培养中,该细胞系仍保持旺盛的再生和分化成苗能力。 相似文献
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一品红离体培养诱导体细胞胚的研究(简报) 总被引:5,自引:0,他引:5
一品红的嫩茎离体培养在附加不同浓度的2,4-D和6-BA的MS培养基上,2.4D浓度为1.0~2.0 mg· L~(-1)的愈伤组织生长量最大。这些愈伤组织转移到MS+6-BA1.0 mg·L~(-1)+NAA0.1 mg·L~(-1)的分化培养基上后,产生了胚性愈伤组织,观察到大量不同时期体细胞胚。带有体细胞胚的愈伤组织进一步转移到无激素 MS培养基上,便发育成小植株。 相似文献
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大叶紫花苜蓿愈伤组织原生质体再生植株 总被引:15,自引:0,他引:15
大叶紫花苜蓿下胚轴诱导的愈伤组织在继代培养基上生长快速,易于分散。继代第12d的愈伤组织原生质体的得率为6.5×107/g鲜重。原生质体培养基为SH基本培养基,含有1.0mg/L2,4-0、0.5mg/LBA、2.0g/LCH、2%蔗糖、6%葡萄糖、5mmol/LMES,培养密度为1.0×105/mL。培养至第12d时的原生质体再生细胞植板率为3.7%。由原生质体形成的小愈伤组织在含2.0mg/L2,4-D的MS固体培养基上大量增殖。增殖的愈伤组织转移至2.0mg/L2-ip+0.1mg/LNAA的B5培养基上,形成体细胞胚并发育成完整植株。 相似文献
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硬粒小麦单倍体原生质体培养及植株再生 总被引:4,自引:1,他引:3
由硬粒小麦(Triticum durum Desf.)×玉米(Zea maysL.)建立的单倍性胚性愈伤组织,在继代培养4 个月后置于含2.0 m g/L2,4-D、3% 蔗糖、200 m g/L水解酪蛋白、146 m g/L谷氨酰胺和300 m g/L天冬氨酸的MS液体培养基中进行悬浮培养,4 个月后形成了生长迅速、由大小不同(0.5 ~5 m m )的愈伤组织块组成的愈伤组织悬浮系。酶解试验表明,2.0% 纤维素酶RS和0.5% 的离析酶效果最好,而液体悬浮培养物和固体培养的愈伤组织(在酶解时用锋利的解剖刀片切成1 m m 左右的小块)都能释放出大量原生质体,但悬浮培养物释放出的原生质体状态较好,胞质更浓厚,用KM8p 培养基以琼脂糖包埋培养方式培养时分裂频率可达5% 左右。由原生质体再生的小愈伤组织经增殖、筛选后可获得胚性愈伤组织,将其转移至分化培养基Ⅰ(0.2 m g/L 2,4-D、1.0 m g/L BAP、0.1 m g/LNAA、3% 蔗糖、200 m g/L 水解酪蛋白、146 m g/L谷氨酰胺和300 m g/L天冬氨酸的MS固体培养基)和Ⅱ(不含2,4-D,其它成分同Ⅰ)上进行分步分化培养可再生出完整植株,分化频率约为20% 相似文献
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将苎麻“浏阳大叶绿”的子叶培养在附加0.5mg/LCPA、0.05mg/LBR的CXW培养基上,可获得淡黄色或浅灰绿色的颗粒状愈伤组织。将愈伤组织转移至附加0.1mg/LCPA、0.05mg/LNAA、1.5mg/LZT的CXW培养基上继代,颗粒状结构非常明显。继续培养在添加2mg/LMet,3g/LYE的上述继代培养基上,可分化出一些相互独立的胚状体,继而发育成为胚状体幼苗。 相似文献
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马尾松成熟合子胚的体细胞胚胎发生和植株再生 总被引:11,自引:0,他引:11
采用马尾松(Pinusm assoniana Lam b.)成熟合子胚为起始外植体,在含2,4-D 10 m g/L,KT和BA 各4 m g/L的DCR培养基上得到胚发生培养物。将白色半透明的愈伤组织(含早期原胚)在含2,4-D1.0 m g/L,KT和BA 各0.4 m g/L的DCR培养基上保持并增殖。在附加9000 m g/L肌醇的DCR高渗培养基上得到粗壮的后期原胚。ABA 和活性炭同时使用能促进子叶胚的形成,最高频率为35.1% 。在无激素培养基上,成熟体细胞胚萌发并进一步形成完整小植株 相似文献
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绞股蓝悬浮细胞的原生质体再生植株 总被引:2,自引:0,他引:2
绞股蓝(Gynostemma pentaphyllum (Thumb)Mak.)是葫芦科多年生草本药用植物,现已得到广泛的开发利用,本文首次报道了绞股蓝悬浮细胞的原生质体再生植株。 相似文献
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白Pian体细胞胚悬浮培养的动力学研究 总被引:1,自引:0,他引:1
白(PiceameyeriRehd.etWils.)是我国特有的云杉属树种,在林业生产和环境绿化中均具有重要地位。其体细胞胚胎发生的研究,一方面可用于优良种质的大规模快速繁殖,为植树造林和园林绿化提供优质苗木;另一方面可作为遗传转化的再生系统,进行树种遗传... 相似文献
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Somatic Embryogenesis and Plantlet Regeneration from Callus of Mature Zygotic Embryos of Masson Pine
Embryogenic cultures were initiated from mature zygotic embryos of masson pine (Pinus massoniana Lamb. ) on DCR medium supplemented with 2, 4-D 10 mg/L, KT and BA each at 4 mg/L. Pale and translucent calli with early stage proembryos were maintained and multiplicated on DCR medium supplemented with 2, 4-D 1.0 mg/L KT and BA each at 0.4 mg/L. Robust late-stage proembryos were obtained when the calli were cultured on DCR medium containing 9000 mg/L myo-inositol. Abscisic acid and activated charcoal promoted the formation of cotyledonary embryos at the highest frequency of 35.1 %. Mature somatic embryos could germinate and develop further into plantlets when they were isolated and cultured on a hormone-free DCR medium. 相似文献
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Formation of Plantlets was achieved when stem apex of Pinellia ternata Brier. Cultured in vitro on MS medium with KT 0. 5 mg/L + NAA 0.2 mg/L (MSI). With petioles of the plantlet as explants callus could be induced after cultured for a week on MS medium with 2, 4-D 2.0 mg/L + KT 0.5 mg/L (MSII). Calli were subcultured once in every month. After 3--4 months a kind of friable calli could be selected, from which the tubercles could be differentiate and the plantlets formed when transfered onto MSI. But before callus differentiation, a lot of roots were formed on callus. The plantlets could be produced directly from the petiole segment. It was found that the stem growing tip was always covered by the leaf primordium and the former leaf primordium was covered by the latter leaf primordium during the differentiation of the apical bud of tubercle. The frenquency of plantlet differentiation from callus and petioles was over 70%. The rate of regeneration of plantlet on liquid static culture was twice as much as that on solid culture. All plantlets grew well after being transfered into the plot. The fresh weight of tuber-plant was 103 % higher than that of control (cultivated plant come from tubers). The alkaloid content of tubers come from tuberplant was 0. 344%, that of control was 0. 203% and 0. 264% for the wild tuber. 相似文献
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Poplar NL-80106 (Populus deltoides×P, simonii) mesophyll protoplasts were isolated from leaves of 30 days-old sterile shoot, with 4 × 107/g fr. wt of protoplast yield after purification. The protoplasts were cultured in KM8p and MS liquid media containing 2 mg/L 2, 4-D, 0. 5 mg/L NAA and 0.5 mg/L KT. Higher plating density and lower osmatic pressure (0.45 mol/L) were proved to be favourable to division of protoplast-derived cells. The first division initiated 5 days after culture, and the division frequency reached 4.5 % on the 10th day. A number'of cell colonies and microcalli was formed in 12 weeks. Using organic nitrates and glucose in protoplast culture medium was beneficial to increase division frequency and plating efficiency. The calli were allowed to grow to 4--6 mm in height with red colour and compact structure on the gelrite-sohdified NLZ1 proliferation medium in 3 weeks and were transferred onto NLF differentiation medium where the frequency of shoot formation could reach 100%. The 3 cm high shoots were then cut off from the callus and rooted on 1/2 MS medium. 相似文献
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太白米组织培养的研究 总被引:2,自引:0,他引:2
太白米地下鳞茎在MS附加不同浓度KT、BA、IAA、NAA、2,4-D的培养基上,可诱导产生愈伤组织,其中在MS附加NAA 0.5mg/L,KT0.1mg/L的培养基上愈伤组织诱导率最高,可达65%,且生长快;培养在MS附加2,4-D 1.0mg/L,KT 0.1mg/L培养基上的鳞茎可经不定根直接发育成新的鳞茎,由此建立了太白米的鳞茎再生体系,鳞茎再生率达2.17倍。 相似文献
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Protoplasts were isolated and cultured from hypocotyl embryogenic callus tissue of Gossypium hirsutum L. cv. "Lumian 6". The highest yields of viable protoplasts were obtained from a vigorous embryogenic callus 7 to 9 d old subcultured on MS medium supplemented with 2 mg/L IAA and 1 mg/L KT using a solution of 1% cellulase Onozuka R-10, 1% pectinase, 0.7 mmol/L KH2PO4, 2.5 mmol/L Ca2+ , and 0.5 mol/L osmoticum (mannitol), at pH 5.8 and at a temperature of 30 ℃. After separation and purification (in 21% sucrose floatation medium), the protoplasts were laid up in a quiet liquid protoplast culture medium containing K3 salts, NT vitamins with 0.1 mg/L 2,4-D, 0.2 mg/L KT and 0.45 mol/L glucose for 10 to 15 min. The protoplasts were fractioned into an upper and a lower layer in the centrifugal tube. Most of the protoplasts in the lower layer were smaller, round and rich in cytoplasts in which contain many granular substances. When this kind of protoplasts were cultured in the thin liquid protoplast culture medium with a density of 1 x l0s to 5 x los protoplasts/mL, the division and the callus formation of the regenerated cells were easily observed. The first divisions occurred in 3 days and small cell clusters could be seen after 2 to 3 weeks in the culture. At this moment, the addition of the protoplast culture medium with decreased osmoticum once or twice is needed for the continuous protoplasts division to form calli. Regenerated calli, 3 to 5 mm in diameter, were transferred in succession on MS medium with 2 mg/L IAA and 1 mg/L KT for the initiation of embryogenesis. The embryoids germinated on the hormonefree MS medium and a number of plantlets were obtained. It seems that using vigorous embryogenic callus and decreasing osmoticum are the two critical factors for plant regeneration of cotton protoplasts. 相似文献
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从甘蔗(Saccharum officinarum L.)嫩叶外植体诱导愈伤组织,经继代培养后,挑选胚性愈伤组织,转入MS3 液体培养基,进行悬浮培养。当培养物分离出小粒状的细胞团,细胞变得小而圆时,用于分离原生质体。原生质体以琼脂糖固化的培养方式培养于MRP1 培养基中。由原生质体再生的愈伤组织有两种类型。挑选粒状、坚实的再生愈伤组织转移到N6 分化培养基上,“新台糖1 号”再生的愈伤组织,在含有KT 0.5 m g/L的培养基中,分化出绿芽并长成完整的植株。而“粤糖57-423”和“US66-56-9”再生的愈伤组织,在加有0.1% 的活性炭的培养基中,前者分化出白化苗,后者分化出根 相似文献
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ACC合成酶基因及其反义基因对西瓜的遗传转化 总被引:21,自引:0,他引:21
以2日龄西瓜(Citrullus lanatus(Thunb.)Mansfeld)无菌苗子叶为外植体,通过与根癌农杆菌(Agrobacterium tumefaciens)进行叶盘共培养建立了西瓜的遗传转化系统。所用根癌农杆菌中含有改建后分别携带嵌合NPTⅡ基因和番茄的ACC合成酶基因及其反义基因的质粒。外植体在MSA培养基(MS盐类、B_5维生素、1.0mg/L BA、0.2mg/L IAA)上预培养3~4d后,与根癌农杆菌共培养4d,随后转移外植体至附加100mg/L卡那霉素、300mg/L头孢菌素的MSA培养基上筛选转化芽。将带芽外植体移入含有100mg/L卡那霉素、300mg/L头孢菌素的伸长培养基(MS 0.2mg/L KT)上进行芽伸长,切取2~3cm高的伸长芽移入生根培养基(1/2MS 0.1mg/L NAA)生根。Southern blot结果证明获得转基因植株,乙烯释放指标表明转入的正义和反义ACC合成酶基因得到不同程度的表达。 相似文献