中国白菜小孢子离体培养和双单倍体植株再生

摘 要: 采用略加修改的NLN培养基（无机大量元素1/2）和甘蓝型油菜小孢子的胚状体诱导方法,培养了5个中国菜用小白菜品种和1个中国大白菜品种的小孢子,进行了双单倍体的诱导。结果表明,不同品种的小孢子产胚量差异很大,在接种的6个基因型中,有5个诱导出胚,诱导成功率83.33％ 。其中的中萁青抗热605产胚量最高,是8.676个胚／10蕾。这些胚在添加了NAA和PP333（多效唑）的B5培养基发育成健壮的小植株。关于6个基因型的小孢子胚胎发生能力,还有待进一步研究。     

Allelopathic Effects of Black Mustard (Brassica nigra) on Germination and Growth of Wild Barley (Hordeum spontaneum)

Black mustard [Brassica nigra (L.) Koch.] contains water‐soluble allelochemicals that inhibit the germination and growth of other species. This characteristic could be used in weed management programmes. Greenhouse and laboratory experiments were conducted to determine the effects on wild barley (Hordeum spontaneum Koch.) germination and seedling growth of (i) preceding crops, (ii) fresh black mustard residue incorporation, and (iii) black mustard leaf, stem, flower and root water extract concentrations. Growth of wild barley, as indicated by plant height and weight, was significantly reduced when grown in soil previously cropped to black mustard compared with that cropped to wild barley. Soil incorporation of fresh black mustard roots and both roots and shoots reduced wild barley germination, plant height and weight when compared with a no‐residue control. In bioassays, black mustard extracts reduced wild barley hypocotyl length, hypocotyl weight, radicle weight, seed germination, and radicle length by as much as 44, 55, 57, 63 and 75 %, respectively, when compared with a water control. Increasing the water extract concentrations from 4 to 20 g per 100 ml of water of all black mustard parts significantly increased the inhibition of wild barley germination, seedling length and weight. Based on 8‐day‐old wild barley radicle length, averaged across all extract concentrations, the degree of toxicity of different black mustard plant parts can be ranked in the following order of inhibition: leaves > flowers > mixture of all plant parts > stems > roots.     

陆地棉原生质体培养与植株再生技术研究

 分离培养陆地棉品种ZDM-3（Gossypium hirsutum L cv ZDM-3）胚性细胞悬浮系的原生质体,通过体细胞胚胎发生成功获得再生植株。试验结果表明,植物生长调节剂组合和碳源对原生质体培养具有重要的影响,添加2,4-D + KT + 1.5%（W/V）葡萄糖+ 1.5%（W/V）麦芽糖的KM8P培养基对于陆地棉原生质体培养的效果较好。激素组合0.46 μmol·L^-1 KT + 0.45 μmol·L^-1  2,4-D诱导的愈伤组织较松软,分化潜力高;胚性愈伤组织的增殖使用0.93 μmol·L^-1 KT + 2.46 μmol·L^-1 IBA的激素组合;1.2 μmol·L^-1 IBA + 1.39 μmol·L^-1 KT有利于体细胞胚胎发生,而MSB-5 + 0.67 μmol·L^-1 KT+2.69 μmol·L^-1 NAA的培养基适合体细胞胚胎萌发和植株再生。此外,愈伤组织诱导宜用葡萄糖作为碳源,而胚性愈伤组织增殖、保存和胚状体的萌发过程宜使用麦芽糖作碳源。     

不同品种橡胶树花药愈伤组织诱导、分化及植株再生的比较

为优化橡胶树花药体细胞胚诱导及植株再生体系,为橡胶树分子育种提供技术支撑,以橡胶树品种‘云研73-477’、‘云研73-46’、‘热研8-79’、‘云研77-4’、‘RRII105’、‘热垦525’的花药为材料,比较不同品种在相同培养条件下愈伤组织诱导、体胚发生能力及植株再生频率。结果表明,6个品种的花药均能脱分化形成愈伤组织,但不同品种间存在显著差异,诱导率最高的是‘RRII105’和‘热垦525’,分别为93.33%、90.33%,最低的是‘云研77-4’,为31.33%;体胚发生能力在不同品种间也有较大差异,‘云研73-477’体胚诱导率最高达76%,‘云研73-46’没有分化出体细胞胚;植株再生频率较高的品种有‘热垦525’和‘云研73-477’,分别为75.38%、54.35%,‘云研73-46’和‘云研77-4’无再生植株。     

Somatic Embryogenesis in Hypocotyl Protoplast Culture of Rapeseed (Brassica napus L.)

By using a system of agirose plating and agarose bead culture, it was possible to induce efficient somatic embryogenesis in protoplast-derived calli of two rapeseed varieties, ‘Ceres’ and ‘Duplo’. Protoplasts were isolated from hypocotyls. For the initial protoplast culture a modified 8P medium was employed containing 2,4D (1.0 mg/l), NAA (0.1 mg/ 1), BAP (0.4 mg/l) and mannitol (7 %). After microcalli were obtained in four weeks, somatic embryos were induced by a two-step method. This involved a modified MS medium containing 2,4D (3.0 mg/l) in the first step and no 2,4D, but BAP (3.0 mg/l) and GA₃ (0.1 mg/l) in the second. This procedure also secured plant regeneration.     

The Influence of Dicamba on Somatic Embryogenesis and Frequency of Plant Regeneration from Cultured Immature Embryos of Wheat (Triticum aestivum L.)

An investigation was made to discover the influence of dicamba on the somatic embryogenesis of winter wheat cultivars-. Immature embryos of Triticum aestivum cv, ‘Sage’, ‘Caribo’ and ‘Kanzler’ were cultured, on modified N6-medium with the addition of 1 mg/13,6 dichlor-2-methoxy benzoe acid (dicamba). The young embryos were placed with the embryo axis on to the medium. Under this condition the scutella of the embryos at different stage of development produced compact calli and embryoids which regenerated plants with a high frequency (70 %) four to: six weeks later. The results suggest that dicamba could be of value in the induction of somatic embryogenesis.     

棉花组织培养高效植株再生体系的建立

通过对影响棉花体细胞胚胎发生和植株再生关键因素的研究,建立了适用于广泛基因型的棉花组织培养高效胚胎发生与植株再生体系。从中棉所12、中棉所19、泗棉3号等20余个主栽品种诱导获得了胚胎发生和植株再生,有效地突破了棉花组织培养植株再生的基因型控制,使占我国棉田面积50%以上的品种均能诱导获得胚胎发生和再生植株。首次诱导获得直接胚胎发生,使棉花组织培养的周期由180d缩短到120d左右,减少了培养过程中变异的发生。该结果的获得必将大大促进植物细胞工程和基因工程在棉花遗传改良上的应用。     

油菜薹茎段髓部原生质体培养再生植株

以油菜品种“蜀杂6号”的薹茎段髓部为材料分离原生质体,产量一般在每g鲜重2×10~6个左右。在每L附加1.5mgBA、0.4mgNAA、1.5mg2,4-D、200mg水解酪蛋白、250mg木糖、0.3M蔗糖和0.1M葡萄糖的Nitsch培养基中,对原生质体进行液体浅层静置培养。2～7天内原生质体出现第一次分裂,分裂频率最高可达49.2％。培养15天后逐渐形成肉眼可见的小愈伤组织。21天时将培养基改换成每L附加2mgBA、0.2mg 2,4-D和3％蔗糖的MS扩增培养基,45天后可形成5mm左右的愈伤组织。大于2mm的愈伤组织可在每L含有1.0mgBA/3.0mgZT和0.1mgNAA的MS琼脂培养基上分化出芽,分化频率可达36.7％。在1/2MS无激素培养基中,1cm左右的分化芽有85.9％生根,形成完整的植株。     

秋水仙碱及热击与低温诱导对油菜小孢子胚状体成苗率的影响

甘蓝型冬油菜F1杂种分离小孢子直接用500 mg/L秋水仙碱处理15 h,在NLN-13培养液中培养,并于32℃和30℃分别热击暗培养3 d和7 d后转入24℃下培养,胚胎发生好,5周后产生大量正常胚体.将这些发育优异的胚体移入固体MS培养基后即进行10 d低温诱导(2℃),常温(24℃)光下培养能很好地萌发并直接、快速地再生植株,其萌发胚率和成苗率     

Synthesis of somatic hybrids (RCBB) by fusing heat-tolerant Raphanus sativus (RR) and Brassica oleracea (CC) with Brassica nigra (BB)

Brassica carinata (BBCC), a potential oilseed crop for dry land agriculture, is sensitive to high temperatures during germination and early stages of growth, which thereby restricts the possibility of using the residual soil moisture available after the rainy season for its cultivation. To overcome this problem, a three‐genome hybrid, RCBB, was synthesized using Raphanus sativus (RR) and Brassica oleracea (CC) as donor sources for the desired heat tolerance. Protoplasts of RC hybrids obtained through sexual crosses between R. sativus (female) and B. oleracea (male) were fused with protoplasts of Brassica nigra (BB) to produce RCBB somatic hybrids. The hybrid colonies regenerated with an average frequency of 7.6%. Twelve out of 36 hybrids grown to maturity were characterized for their nuclear and organelle genomes. While all the hybrids showed the presence of B. nigra chloroplasts, 10 of the hybrids showed B. nigra‐specific mitochondria and two had Raphanus‐spedfic mitochondria. The somatic hybrids could be backcrossed to B. carinata.     

Anther Culture in Rye: Improved Plant Regeneration Using Modified MS-Media

The use of androgenetic doubled haploids in rye breeding is still limited by low regeneration rates. In this study we tested the influence of two modified MS-media on the number of reacting anthers and on plant regeneration. Picloram and 2,4-D were used as auxin components in the induction media. The highest induction rates (reacting anthers per 100 cultivated anthers) and the highest regeneration rates (number of green plants per 100 cultivated anthers) were reached on a modified MS-medium containing Picloram as auxin source, regardless of the donor plant material. The comparison of the results obtained with the single cross ‘SC 35’ which contains genes of Secale vavilovii and the true Secale cereale donor forms clearly show the genotypic influence of donor plant material on induction and regeneration ability.     

Assessment of transgene escape from Brassica rapa (B. campestris) into B. nigra or Sinapis arvensis

The possibility of gene transfer between Brassica rapa and the two weedy species B. nigra and Sinapis arvensis was evaluated with the special concern on transgene escape from B. rapa to these two weedy species. B. rapa cultivar Tobin was reciprocally crossed to five and four strains of B. nigra and S. arvensis, respectively, using controlled cross. A single interspecific hybrid was obtained from the cross B. rapa×B. nigra, but no other cross was successful. The fertility of this hybrid on open pollination, selfing and backcrosses was investigated. The data of the present study and the information available to date indicate that gene transfer between B. rapa and B. nigra is possible. The chance of transgene escape from B. rapa to B. nigra depends essentially on whether natural cross can occur between these two species. Gene transfer between B. rapa and S. arvensis is at the most difficult, whereas trans-gene escape directly from B. rapa to S. arvensis appears very unlikely.     

大白菜无菌苗子叶组织培养再生植株

选用8个大白菜亲本材料进行离体培养试验,探讨了植物激素、品种、苗龄、取材部位及外植体放置方式对大白菜子叶不定芽再生的影响。研究结果表明,大白菜子叶再生培养基为MS附加6-BA 2mg／L,NAA 0.5mg／L和AgNO3 5mg／L;生根培养基为1／2MS附加NAA 0．2mg／L。再生植株移栽时,最好在春季,成活率可达90％以上。     

黑糯米成熟胚愈伤组织培养及植株再生研究

对贵州惠水黑糯、黑糯141成熟胚愈伤组织培养及再生条件的研究结果表明,分步消毒可以减少外植体的污染率,在附加2,4-D 2 mg/L的NBD 1培养基中,成熟胚愈伤组织诱导率较高,分别为93.84%、90.52%。愈伤组织的继代培养是分化前必不可少的过程,适当干燥处理及合适的激素配比能够提高愈伤组织分化率,贵州惠水黑糯分化率为95.18%,黑糯141分化率为89.38%。2个黑糯米品种在附加NAA 0.5 mg/L的生根培养基中均能正常生根。     

A Simple Method of Inducing Somatic Embryogenesis in Brassica juncea (L.) Czern & Coss.

A single-step method for the induction and development of somatic embryoids from hypocotyls explains of Brassica juncea is reported. On modified MS medium containing 2 % sucrose, 0.25 mgl ¹ 2,4-D, 0.5 mgl ¹ each of NAA and BaP-R, each explant calluses at both of and at its best, 31% of explants produce embryoids. In the variety RLM-198, the number of embryoids ranges from 8–21 per culture. Each embryoid, upon proliferation, developed up to the 25 shoots. The method is rapid; the time La ken from inoculation to the development of intact plantlets is 8–10 weeks. Regenerated plants have flowered normally and have set seed. The system can profitably be used for in vitro mutant selection and early bulking in mustard.     

水仙离体培养及植株再生的研究进展

摘 要：水仙离体培养及其再生技术在水仙品种改良、种质资源保存与繁殖、遗传转化等方面发挥着重要作用。本文综述了20世纪70年代以来水仙离体培养及植株再生的研究进展,包括水仙营养器官培养、生殖器官培养、体细胞培养等方面的应用,对水仙离体培养过程中存在的一些问题进行了探讨并对今后的研究方向作下展望。     

The Effect of Sugars and Growth Regulators on Embryoid Formation and Plant Regeneration from Barley Anther Culture

The influence of carbon source and growth regulator composition in induction medium on anther culture response was investigated using spring barley genotypes. Anthers were cultured on BAC3, Ficoll-containing medium, supplemented with one of the following carbohydrates: sucrose, maltose, cellobiosc and melibiose (6 % w/v). The use of either maltose or cellobiose resulted in a significantly higher anther response, calli and/or embryoid production and green plant regeneration compared to the incubation of anthers on a medium containing sucrose. Contrary to these results, the replacement of sucrose by melibiose in BAO medium, drastically reduced the efficiency of anther culture. As an average for the three genotypes tested, the frequency of green plants per 100 anthers plated was 9- to 22-fold higher on medium supplemented with sucrose or cellobiose than on medium containing melibiose as a sole carbohydrate. Among the growth regulators tested, the combination of auxin NAA (2 mg/l) and cytokinin BAP (1 mg/1) performed much better than the employment of auxin 2,4-D combined either with zeatin riboside or BAP as cytokinins. The beneficial effect of medium supplemented with NAA and BAP was associated with better embryoid formation compared to the other growth regulator combinations tested. The hormone-free combination gave a similar anther response and production of calli as any medium supplemented with growth regulators, but the regeneration capacity of calli produced on hormone-free medium was much lower, resulting in the drastic reduction of the number of both total and green regenerants.     

Plant Regeneration Through Anther Culture of Three Wild Species of Hordeum (H. murinum, H. marinum and H. bulbosum)

Plant regeneration was achieved through anther culture of three wild species of Hordeum (H. murinum, H. marinum and H, bulbosum). Calli or embryoids were formed from microspores in anthers cultured on a medium containing 6-benzylammopurine (BAP) and ficoll. These calli or embryoids regenerated green or albino shoots and roots after transfer to regeneration media. Green plantlets which developed on regeneration media were transferred to soil where they showed further growth.     

Effect of genotype,environment and carbohydrate on anther culture response in head cabbage (Brassica oleracea L. convar. capitata (L.) Alef.)

Summary The effect of genotype, growing conditions for donor plants and type and concentration of carbohydrate in the culture medium was investigated for anther culture of head cabbage (white cabbage, savoy cabbage, pointed-headed cabbage). Strong genotypic effects on embryo formation from the cultured anthers were shown as well as superior embryo formation from anthers of field grown donor plants compared to plants grown in the greenhouse. When comparing 7, 10 and 13% sucrose in the medium, embryo response increased with increasing sucrose concentration. With maltose, which was generally inferior to sucrose as carbohydrate source for anther culture, the embryo response did not increase with maltose concentration above 10 per cent.