In-vitro Fertilization of Ovules of Some Species of Brassicaceae*

The method of in-vitro fertilization of ovules can be successfully applied to various species of Brassincaceae. Mature embryos and plants were obtained after in-vitro pollinating the ovules of Arabis caucasica, Brassica napus, B. oleracea var. sabellica (kale), B. oleracea var. italica (broccoli), Diplotaxis tenuifolia, Moricandia arvensis and Sisymbrium Loeselli. In the case of Sinapis alba fertilization and embryo development did not occur. The same method has been successfully used for obtaining hybrid immature embryos at different stages of development from crosses between B. napus X D. tenuifolia, B. napus X M. arvensis, B. oleracea var. italica X D. tenuifolia, D. tenuifolia x B. napus, D. tenuifolia. X M. arvensis and D. tenuifolia X S. Loeselli. The present findings show that in-vitro pollination of ovules of various species of Brassicaceae makes it possible to (a) perform the whole process of fertilization and embryogenesis; and (b) obtain intergeneric hybrid embryos.     

ABA对花生胚离体发育的调节

实验结果表明，ＡＢＡ使花生离休胚处于胚性状态、抑制早萌并继续发育，ＡＢＡ的作用与其浓度有关。ＡＢＡ促进贮藏蛋白的合成，这种调控作用与胚的发育阶段有关。ＡＢＡ也促进了花生胚热稳定蛋白的合成和累积．     

鸡新城疫、传染性支气管炎、传染性法氏囊病三联活疫苗的研究

鸡新城疫(ND) La Sota、传染性支气管炎(IB) H120和传染性法氏囊病(IBD) B87弱毒株适当稀释后等量混合,经10日龄SPF鸡胚同胚接种联合培养,收获含毒鸡胚液和胎儿混合制成ND、IB、IBD三联活疫苗。接种7~14日龄SPF雏鸡7 d产生抗体,14~21d达到高峰,免疫期70 d以上,与单苗同步免疫并攻击强毒,试验结果无显著性差异。以10个使用剂量免疫SPF雏鸡,无ND、IB、IBD临床症状和剖检病变,其安全性和效力检验均达到相应单苗的标准要求。     

AE油佐剂灭活疫苗对鸡胚和仔鸡的保护力试验

三批禽脑脊髓炎(AE)油佐剂灭活疫苗免疫开产前种鸡,种鸡免疫2~3个月后所产种蛋对禽脑脊髓炎病毒(AEV)鸡胚易感性试验的受保护率达100%,而同期所产种蛋孵出的仔代雏鸡,在出雏后3周内对AEV VR株攻毒的受保护率亦达100%; 种鸡免疫6~7个月后所产种蛋对AEV VR鸡胚易感性试验的受保护率达75%~80%,而同期所产种蛋孵出的仔代雏鸡,在出雏后3周内对AEVVR株攻毒的受保护率达70%~90%。试验结果表明,AE灭活疫苗AEV鸡胚易感性试验的鸡胚保护率与同期雏鸡对AEV VR肌肉途径攻毒的保护率基本相同。     

The Effectiveness of Vernalization of Immature Embryos of Winter Wheat var. Grana as Related to Age and Exogenous Phytohormones

Starting from the 10th day after pollination, immature embryos of winter wheat var. Grana were isolated and then vernalized for 4 to 8 weeks on Murashige and Skoog nutrients containing BAP (0.5 or 2 mg/dm³), NAA (0.5 or 2 mg/dm³), or GA₃ (5 or 20 mg/dm³). Vernalized seedlings were cultivated in a greenhouse and the number of days to the heading of individual plants as well as the percentage of plants capable of generative development were estimated. The lower limit of size for 50 % survival of embryos strongly depended on the phytohormone used: from 0.9 mm in control, 1.1 mm in nutrient containing BAP, 1.2 mm for NAA, up to 1.7 mm in nutrient with GA₃. Exogenous GA₃ was lethal for embryos younger than 18 days but induced elongation of older embryos. Embryos isolated 2.5 to 4 weeks after pollination showed minimal requirements for the length of vernalization. BAP increased the percentage of heading plants originated from the youngest embryos. GA₃ improved partial vernalization, strongly increasing the percentage of heading plants, but did not change the time from the end of vernalization to heading. It has been postulated that GA₃ increases number of plants capable of overcoming the threshold of induction of generative development but does not accelerate the flowering process. Hormonized plants showed no deformation of generative development. As the effectiveness of vernalization increased, the heading of plants was faster but they were shorter, forming spikes with a smaller number of spikelets and producing fever lateral shoots. The very young embryos probably have in reserve sufficient amounts of auxins and gibberellins and therefore exogenous GA₃ decreases their viability or even exerts a deleterious effect. However, as the embryos' ageing, gibberellin starvation develops. This being especially pronounced during vernalization. The de novo synthesis or activation of gibberellins takes place during the second stage of vernalization. This is why exogenous hormone improves the effectiveness of partial vernalization, though it is not possible to substitute by gibberellins the vernalization requirements of immature embryos.     

RNA Synthesis in Porcine Blastomere Nuclei Introduced into in Vitro Matured Ooplasm

The objective was to investigate the RNA synthesis in porcine blastomere nuclei upon transplantation into in vitro matured enucleated oocytes. Nuclei from 2- to 8-cell porcine embryos were introduced into the ooplasm of in vitro matured and enucleated porcine oocytes by electrofusion, and the resultant reconstructed embryos were cultured in vitro. Before fusion or at different intervals after this event embryos were incubated with [³H]-uridine, fixed, and histologically processed for autoradiography in order to detect RNA synthesis. About two thirds of the embryos were considered to depict normal development. All blastomeres displayed pronounced RNA synthesis before fusion, at 3 and 9 h after fusion the synthesis decreased or ceased, and at 24-49 h some embryos resumed synthesis at the 1- to 2-cell stage.     

牡丹‘凤丹’体细胞胚发生技术

为建立牡丹‘凤丹’体细胞胚发生培养体系,以牡丹‘凤丹’的合子胚、胚轴和子叶为外植体进行离体培养,研究了种胚发育时期不同质量浓度蔗糖处理及不同的培养基对牡丹体细胞胚发生的影响,结果表明,花后110 d是体细胞胚直接诱导合适的发育时期,诱导率为33.00%,3种外植体中,种胚的诱导率最高;利用质量浓度为90 g.L-1的蔗糖溶液处理种胚2 h,体细胞胚诱导效果最好,在MS+2,4-D 2.0 mg.L-1+6-BA 2.0 mg.L-1培养基上获得较高的体细胞胚,诱导率为38.33%。     

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。     

陕西关中中部灌区12个小麦品种幼胚脱分化特性研究

[目的]对陕西关中中部灌区推广的12个小麦品种幼胚脱分化特性进行研究,以筛选出优良转化受体的基因型。[方法]供试小麦品种为综合农艺性状优良的小偃22、西农9871、西农979、阎麦8911、西农889、西农2611、陕麦159、陕农138、西农2000、西农3517、小偃216和西农538。培养基A:MS基本培养基+2,4-D 2.0 mg/L+6-BA 0.5 mg/L+蔗糖30 g/L+琼脂4 g/L;培养基B:MS基本培养基+2,4-D 2.0 mg/L+6-BA 0.5 mg/L+AgNO32.5 mg/L+蔗糖30 g/L+琼脂4 g/L。培养温度25～26℃,暗培养,28～35 d继代培养1次。[结果]不同小麦品种之间愈伤组织的形成时间、愈伤组织诱导率、胚芽率和胚性愈伤组织诱导率明显不同,且在2种培养基间的波动趋势具有一致性;培养基中附加的AgNO3可以减轻胚发芽的发生概率而促进胚性愈伤组织产生;12个品种中,小偃22、陕农138和小偃216表现出较好的脱分化综合性状,可用于小麦的基因转化研究。[结论]该研究可为建立高效稳定的小麦植株再生体系奠定基础。     

不同菜用大豆品种的胚胎发生与再生

以8个菜用大豆品种的未成熟子叶为外植体,在MS(附加高浓度2,4-D)培养基诱导愈伤组织形成、体细胞胚胎发生与再生植株.结果表明:八个菜用大豆品种的未成熟子叶均可诱导产生愈伤组织与体细胞胚胎发生,出愈率为14.71%～100%.体细胞胚胎发生率变化在0～36.76%之间,绿子叶菜用大豆没有形成体细胞胚.体细胞胚可正常萌发产生小植株,移栽成活结实.菜用大豆的出愈率和体细胞胚胎发生率与基因型有关.