水稻“双－3”多胚发生的研究

叶秀,陈泽濂,黎垣庆 　　水稻ＭＩＶ（双－３、籼稻）传粉后可以有多个花粉管同时进入胚囊．大多数胚囊的合子发育为一个正常的胚,但是有少数合子胚发生裂生并分化形成双胚芽和一胚根．有些胚囊的助细胞和卵细胞同时受精后,分别发育为助细胞胚和合子胚;有些胚囊中的反足细胞团可直接发育为胚．可见“双－３”水稻除有正常合子胚外还存在助细胞胚和反足细胞匹的多胚现象．     

小鼠分离胚的培养和移植

将小鼠2—4细胞胚分离成为1/2半胚或2/4半胚后,在体外条件下进行培养,选择发育为桑椹胚、囊胚期的胚胎移植给假孕雌鼠。结果是,分离胚的囊胚发育率在Whitten和BMOC-Ⅲ中分别为88.1％和81.6％,1/2半胚和2/4半胚的发育率分别为89.0％和94.6％(p<0.05),F_1代和ICR1/2半胚的发育率分别为91.7％和49.2％(p<0.01)。在移植1/2半胚和2/4半胚的雌鼠中分别有3只和1只妊娠并产仔鼠2只和1只;在以1/2半胚,去透明带的整胚和保留透明带的整胚为对照移植的三个处理组中其雌鼠的妊娠率和产仔率分别为8.3％和2.8％,30.0％和10.8％,60.0％和36.7％,各处理组间均有显著差异(p<0.01)。     

水稻“双－3”多胚发生的研究

水稻ＭＩＶ（双－３、籼稻）传粉后可以有多个花粉管同时进入胚囊．大多数胚囊的合子发育为一个正常的胚,但是有少数合子胚发生裂生并分化形成双胚芽和一胚根．有些胚囊的助细胞和卵细胞同时受精后,分别发育为助细胞胚和合子胚;有些胚囊中的反足细胞团可直接发育为胚．可见“双－３”水稻除有正常合子胚外还存在助细胞胚和反足细胞匹的多胚现象．     

胡萝卜(Daucus carota L.)胚性细胞蛋白的分离研究

应用IEF/SDS-PAGE双向电泳技术,比较了胡萝卜胚性细胞、非胚性细胞和不同发育时期的胚状体中可溶性蛋白的双向电泳图谱,结果发现在胚性细胞中特异存在的胚性细胞蛋白在不同发育时期的胚状体中也存在,但在非胚性细胞中不存在。因此,推测体细胞胚胎发生所需的一些基因在胚性细胞中就早已表达了。我们还成功地分离和测定了ECP 45-2 N-末端和中央部分氨基酸序列。与已知氨基酸序列相比,ECP 45-2部分氨基酸序列与ECP 45-1 部分氨基酸序列具有较高比例的同源性。因此, ECP 45-1和45-2可能属于同一因基家族。     

多胚水稻ApⅢ（双13）的胚胎学观察

对多胚水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）ＡｐⅢ的大量成熟颖果、人工萌发的幼苗和开花后３～５ ｄ 的幼嫩颖果进行的整体解剖和显微制片观察表明：ＡｐⅢ的５０００粒成熟颖果中,８９．０％ 含单胚单苗,８．９％ 和１．２％分别含双胚双苗和三胚三苗;７００多粒幼嫩颖果中,９０．０％ ～９５．０％ 含单胚,５．０％ ～７．０％ 含双胚。因制片的数目有限,未见到含三胚的;在含单胚和多胚颖果中,胚均位于同一胚囊的珠孔端,未见到胚囊以外存在不定胚。根据上述结果,似可以认为ＡｐⅢ单粒颖果的双胚和三胚是由同一胚囊内的卵细胞和１或２个助细胞受精或不受精发育而来的     

红肉小果型番木瓜品种'美中红'体胚的诱导

用红肉小果型番木瓜品种‘美中红’为外植体,探讨不同成熟度的幼胚、不同浓度2,4-D和培养条件对其体胚的诱导以及体胚形成过程的结果表明:以子叶和内外种皮都为白色且个体较大的番木瓜幼胚(90~120d)在含10mg·L-12,4-D的培养基中和黑暗条件下诱导愈伤组织的效果最佳,愈伤组织的诱导率随着2,4-D浓度的增加而增加。番木瓜愈伤组织最先发生于形态学上的胚根下端,体胚多发生于形态学上的胚芽上端。     

胚龄和激素对小麦幼胚组织培养的影响

以扬麦158为试验材料,通过田间取样室内培养的方法研究了胚龄和激素对小麦幼胚组织培养的作用。结果表明,幼胚组织培养最适宜的胚龄为14—16d;适宜的2,4-D浓度为1．5-2．5mg/L;适宜的IAA浓度为2．0-3．0mg/L;适宜的6-BA浓度为0．1-0．8mg/L;适宜的KT浓度为0．5—1．5mg/L。因此,胚龄和激素对于小麦幼胚组织培养具有明显的调节作用．在组织培养实践中,充分认识和综合协调这些因素对小麦幼胚组织培养的作用,可以提高组织培养效率,使其更加有利于生物学研究、遗传转化和快速育种等工作。     

沙打旺悬浮培养细胞原生质体的体细胞胚胎发生再生植株(英文)

以继代培养5—24代的沙打旺(Astra-galus adsurgens Pall.)胚性悬浮系为材料,从生长4d的细胞培养物中可游离出大量有活力的原生质体。细胞预质壁分离或低温预处理对原生质体得率和活力没有影响,但可提高原生质体培养的植板率。预处理的原生质体培养在NH_4NO_3浓度降至2.5mmol/L,并附加0.5mg/L NAA、1.0mg/L 2,4-D、0.7mg/L BA和0.4mol/L葡萄糖的KM8P培养基中,经持续分裂形成细胞克隆,植板率高达16%—20%。细胞克隆在附加1.0mg/L 2,4-D和0.5mg/L BA的MS培养基中增殖后,转至含0.1mg/L NAA和1.0mg/L BA的MS分化培养基中可形成体细胞胚,平均体细胞胚发生频率约为40%,每个克隆产生20—40个体细胞胚。在无激素的1/2MS培养基中,体细胞胚发育成形态正常和染色体数稳定的小植株。     

沙打旺胚性原生质体培养优化及高频再生植株

外植体类型和光照条件决定沙打旺胚性愈伤组织的形成。用生长10d的胚性愈伤组织可分离到1.2×10⁶个/g(原生质体/细胞),活力超过80%。当原生质体以1.0×10⁵/mL的植板密度培养在含0.6%琼脂糖附加1.5mg/L 2,4-D、0.5mg/L BA和0.5mol/L葡萄糖的培养基(无机盐降为1/4)中,植板率为16.8%。条件培养基显著促进原生质体的生长发育。长大的细胞克隆经2周4℃低温处理后转到含0.1mg/L NAA和1.0mg/L BA分化培养基上,体细胞胚胎发生频率高达70%,每克细胞产生的体细胞胚数在200个以上。成熟的体细胞胚转到无激素的1/2MS培养基中即分化成苗,再生植株为正常的二倍体。     

Somatic embryogenesis and plant regeneration from protoplasts of asparagus (Asparagus officinalis L.)

Protoplasts were isolated from embryogenic calli of Asparagus officinalis L. cv. Mary Washington and cultured in 1/2 MS medium with 1 mg/l NAA, 0.5 mg/l zeatin, 1 g/l L-glutamine, 0.6 M glucose and 0.1% Gellan Gum. Protoplasts started to divide after 3–4 d of culture and formed visible colonies after 30 d of culture. The percentage of colony formation (plating efficiency) was 7.2%. The colonies were then transferred onto Gellan Gum-solidified MS medium containing 1 mg/l 2,4-D and 3% sucrose for further growth. Somatic embryos were induced from all colonies of 0.5–1.0 mm size after transferring to 1/2 MS medium lacking growth regulators. After treating these somatic embryos (1–3 mm) in distilled water for a week, 30–40% of them germinated normally and grew into plantlets 20–30 d after transplanting on 1/2 MS medium containing 1 mg/l IBA, 1 mg/l GA₃ and 1% sucrose. These protoplast-derived plants were diploid with 20 chromosomes.Abbreviations BA 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid - GA₃ gibberellic acid - IBA indole-3-butyric acid - MS Murashige and Skoog (1962)     

Plant Regeneration from Protoplasts of Coriandrum sativum

Calli produced from stem segments of seedling of Coriandrum satwum which were cultured on MS agar medium containing NAA 1.0mg/L. The embryogenic cell colony suspension was estabilished on MS liquid medium containing NAA 1.0mg/L%2,4-D 0.2mg/L+BA 0.5 mg/L. The cell suspension culture was used for protoplast preparation. Protoplasts were obtained in the enzyme mixture containing 2.0% Onozuka R-10, 1.0% pectinase, 0.5% snailase, 0.5% dextran sulfate potassium Salt, 0.6mol/L mannital CPW solution at pH 5.8 and 25℃. Cultured in a KM8P liquid medium containing NAA 1.0mg/L+2,4-D 0.2mg/L+6-BA 0.5 mg/L, glucose 0.4mol/L and CM 20mi/L; the protoplasts entered the stage of derision after three days, cell clusters formed in 10 days and calli formed after about 50 days. When the calli were transferred to MS agar medium containing many growth substances, they differentiated into embryoids, and then developed into plantlet with many green leaves and roots on the 1/2 MS agar medium.     

龙牙匆木幼芽体细胞胚胎发生和植株再生（简报）

Explants excised from the young shoots of Aralia elata (Miq.) Seem. were cultured on MS media. Calli were induced from the explants on MS medium supplemented with 0.5 mg/L 2, 4-D, 0.5 mg/L BA and 0.5 mg/L NAA. Then these calli were transferred onto the MS medium containing 2.0 mg/L 2,4-D + 0.5 mg/L BA + 0.5 mg/L NAA and 0.2% activated charcoal. Under these conditions the somatic embryoids were observed and regenerated plants were obtained from somatic embryogenesis. Then, a experimental system with stability and high regenerating efficiency has been set up for the propagation of the young plants, the cell breeding technology and the control of somatic embryogenesis of Aralia elata (Miq.).     

三叶半夏悬浮培养下的体细胞胚胎发生及植株再生

用三叶半夏幼嫩叶片诱导产生的胚性愈伤组织建立了胚性细胞悬浮系,研究了悬浮培养下体细胞胚胎的发生及植株的再生。结果表明,胚性悬浮细胞在附加1．0mg／L2,4-D、0．2 mg／LBA和300mg／L LH的MS液体培养基中产生大量的球形胚,转入液体分化培养基(MS 0．1 mg／LNAA 0．2 mg／L BA 300 mg／L LH)中进一步发育成心形胚、鱼雷形胚和子叶形胚。收集成熟胚转移到MS固体分化培养基上培养获得了再生植株。另外还观察到某些成熟胚上产生了许多次生胚。     

Study on Tissue and Protoplast Culture of Wild Cotton (Gossypium davidsonii)

This paper deals with the study on the condition of callus formation, embryogenesis, organogenesis, plant regeneration and protoplast culture of wild cotton (G. davidsonii) Callus cultures derived from several organs such as root, stem, leaf, cotyledon and hypocotyl. The results obtained in these cultures showed that the modified MS medium containing 2,4-D 1.0+KT 0.1; 2,4-D 0.1+KT 0.01; NAA (IAA) 2.0+KT 0.1 and NAA (IAA) 1.0+KT 0.1 mg/L were favorable to callus formation. Modified MS medium containing 2,4-D was suitable for initiated callus of G. davidsonii Besides, suspension cultures from callus of G. davidsonii were saccessfully initiated. Optimum concentration of 6BA (or ZT, or 2ip) and NAA (IAA) was for shooting, somatic embryo or leaf formation. Plantlets regenerated from somatic embryo at lower concentration of 6BA, or ZT, or 2ip. As to protoplast culture of this species, the age and physiological condition of callus or suspension cells and concentration of enzymes used for protoplast isolation affected the yield and survival of protoplasts. Protoplast of this species cultured in modified MS medium containing 2,4-D 0.5+NAA 0.5+ZT 0.1–0.2 mg/L. and divied after 3–4 days. The rate of division was 3--4% and cell cluster formed after 14 days, then these cells died.     

新疆天山雪莲体胚诱导与分化研究

以新疆天山雪莲的叶片为外植体,分别用不同配方培养基诱导愈伤组织,后进行体胚诱导和分化培养形成再生雪莲植株.结果表明,诱导愈伤组织的最适培养基为MS 2,4-D 0.5 mg/L BA 1.5 mg/L,诱导率可达到100%;愈伤组织转移至MS 2,4-D 0.5 mg/L BA 1.5 mg/L培养基进行继代培养,增殖后的愈伤组织转移到MS 2,4-D 0.2 mg/L的液体培养基后成功诱导出雪莲体胚,出胚率达40%;将体胚接至MS ABA 0.5 mg/L培养基后,结果分化生长出大量的再生雪莲幼苗.     

从石刁柏原生质体培养再生植株

石刁柏,又名芦笋(Asparagus officinalisL.)是百合科天门冬属植物。其栽培品种含有丰富的维生素类及蛋白质。同时,石刁柏对于某些疾病有一定的药效,因此它已成为人们所喜爱的一种高级营养蔬菜。国外已有不少关于石刁柏试管苗繁殖的报告,但至今只有Bui Dang Ha等从石刁柏枝状叶分离的原生质体得到愈伤组织,并由此愈伤组织诱导获得了再生植株。此后,未见在石刁柏的原生质体培养方面再有新的工作。在本文中,我们利     

Plant regeneration from callus protoplasts of the forage legume Astragalus adsurgens Pall.

A reproducible release of viable protoplasts was obtained from friable calli of Astragalus adsurgens. Protoplasts underwent sustained divisions and formed cell colonies when cultured in either liquid or agarose-solidified Kao and Michayluk (1975) protoplast medium (KM8P) supplemented with 1.5 mg/l 2,4-D, 0.5 mg/l BA and 0.5 M glucose. Compared to liquid culture, agarose bead culture improved division frequency effectively, the two culture systems showing a plating efficiency of 0.8±0.5% and 6.5±0.7%, respectively. Upon transfer to Murashige and Skoog (1962) medium (MS) with 1–2 mg/l BA, alone or in combination with NAA or 2,4-D at 0.1 mg/l, the protoplast-derived calli produced complete plantlets through somatic embryogenesis. The maximum percentage of calli producing somatic embryos (52.5± 2.2%) occurred on MS medium containing 0.1 mg/l NAA and 1 mg/l BA, whereas the maximum number of calli regenerating plantlets (64.7±6.2) was obtained on MS medium with 0.1 mg/l NAA and 2 mg/l BA. Received: 25 April 1997 / Revision received: August 1997 / Accepted: 2 September 1997