Cytological and Isozyme Evaluation of Tall Fescue × Italian Ryegrass Hybrids

Development of tall fescue (Festuca arundinacea Schreb., 2n = 6x - 42) × Italian rye-grass (Lolium multiflorum Lam., 2n = 2 ×= 14) hybrids would enhance efforts to improve the quality of tall fescue. Two ‘Kentucky 31’ tall fescue בLemtal Italian ryegrass hybrids were obtained via embryo rescue on MS media containing casein hydrolysate, ascorbic acid and sucrose. Chromosome pairing at metaphase I had an average of more than 12 bivalents per cell. Since Festuca-Lolium pairing can account only for seven of the paired chromosomes, intergenomic as well as interspecific chromosome pairing is indicated. There was no cytoplasmic effect on chromosome pairing. To determine if enzymes could be used as genetic markers for distinguishing hybrids from self-contaminants in crosses, zymograms of PGI, 6-PGD, MDH, GOT and ACPH were obtained from parents and hybrids using starch gel etectrophoresis. PGI, 6-PGD and MDH had fewer bands in the diploid ryegrass, as compared with the hexaploid tall fescue and the tetraploid hybrid.     

荔枝胚性悬浮细胞系的快速建立及其体胚植株的再生

荔枝幼胚诱导的胚性培养物在低糖条件下连续继代4~6次左右,可筛选到颗粒细小、不含原胚的松散型胚性愈伤组织;以这种松散的胚性愈伤组织作为起始材料,在附加2,4-D 2mg/L或2,4-D 2mg/L、KT1 mg/L、AgNO3 5mg/L的MS液体启动培养基上振荡培养（100~120 r/min）10~14 d,即可建立起分散性良好的胚性悬浮细胞系。采用激素减半的2种启动培养基交替继代培养或周期性固体－液体轮回培养,可以长期保持胚性悬浮细胞系。荔枝胚性悬浮细胞在附加NAA 0.1 mg/L、KT 或Ze 5 mg/L、肌醇100 mg/L、蔗糖50g/L、琼脂10g/L的MS固体培养基上诱导体胚,25~40d后可形成大量胚状体,诱导体胚数量达10,000个/g FW以上。经过成熟培养后,正常的体胚75%以上萌发再生完整植株。     

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。     

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。     

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。     

Somatic Embryogenesis and Plant Regeneration from Tritordeum

Regeneration of plants by somatic embryogenesis from immature embryos of hexaploid tritordeum (AABBH^chH^ch, amphiploid Hordeum chilense×Triticum turgidum conv. durum) and durum wheat (Triticum tergidum) was induced on MS medium supplemented with different 2.4-D concentrations. Well-defined embryoids were formed with a high frequency on the scutellar callus from 1 or 2 weeks onwards and plantlets were developed from them. In the best cases from one single explant more than 100 plants could be obtained. Plants were also regenerated by somatic embryogenesis from inflorescences of Hordeum chilense×Triticum turgiditm conv. durum hybrid and its respective hexa-amphiploid. With regard to callus induction and regenerative ability, evident differences between hexa- and octoploid (H. chilense×T. aestivum) tritordeum were found, the latter showing a very low response.     

大蕉幼雄花易碎胚性愈伤组织长期继代培养及其植株再生

大蕉未成熟雄花接种到胚性愈伤组织诱导培养基中,4～5个月后可诱导出胚性愈伤组织,并可在继代培养基上长期增殖。这些继代培养了6年多的松软易碎的胚性愈伤组织转移到含有0.2 mg/L 6-BA的分化培养基中,可诱导出芽,得到了53株再生植株,这些再生植株可进一步扩大繁殖。组织学切片证明长期继代培养的愈伤组织维持了胚性的状态。     

野生阿宽蕉胚性细胞悬浮系原生质体分离及植株再生

本研究以野生阿宽蕉(Musa itinerans Cheesman)未成熟种子诱导的胚性细胞悬浮系为材料,对其原生质体的分离和植株再生进行研究。结果表明:胚性悬浮细胞在酶组合:3.0%纤维素酶R-10、1.0%离析酶R-10、0.2%果胶酶Y-23、1.52%KCl、0.78%CaCl2、0.01%MES和11%甘露醇中,酶解8h,原生质体的产量和活力达到最大值,分别为19.46×106个/mL和92.17%。采用看护培养法对原生质体培养60d后,可获得大量增生的小细胞团,细胞团悬浮培养15d后转至体胚诱导培养基上培养约30d,其体胚萌发率为45.97%,然后转至生根培养基上培养35d,其再生植株率为54.76%。     

甘薯胚性细胞悬浮培养系的建立

地甘薯胚性细胞悬浮增减系的进行了研究。将１２个基因的长约０．５ｍｍ的茎尖培养在含有０．２ｍｇ／Ｌ或２．０ｍｇ／Ｌ２，４－Ｄ的ＭＳ培养基上，形成了胚性愈伤组织。胚性愈伤组织的形成率因基因型和２，４－Ｄ深度不同而很大差异，为０－７５．７％。一方面，将胚性愈伤组织继续增减在含有２，４－Ｄ的ＭＳ培养基上，它们形成了处于各发育时期的体细胞胚。将具有体细胞胚的胚性愈伤组织转移到ＭＳ基本培养基上，体细胞胚发育成     

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。     

苎麻体细胞植株再生体系优化研究

旨在探索苎麻体细胞植株再生体系的优化方案,为苎麻组织培养及品种改良提供理论与技术依据。本试验以‘湘苎3号’为试材,优化苎麻愈伤诱导植株再生体系,再将此优化的再生体系应用于其他4个种质材料,得出5个种质的愈伤诱导率、芽诱导率、再生苗生根率,来比较不同基因型之间的体细胞植株再生率差异。结果表明‘,湘苎3号’最佳愈伤诱导培养基为附加2,4-D (0.1 mg/L)和6-BA (2.5 mg/L)的MS培养基;‘湘苎3号’、‘咸丰大叶绿’、‘A’、‘B’、‘中苎1号’的愈伤诱导率分别是:83.3%、80.0%、76.7%、70.0%、53.3%。‘湘苎3号’最佳芽诱导培养基为附加TDZ (0.5 mg/L)、2,4-D (0.02 mg/L)和IAA (0.03 mg/L)的1/2MS培养基,‘湘苎3号’、‘咸丰大叶绿’、‘A’、‘B’、‘中苎1号’的芽诱导率分别是:28.4%、21.2%、12.8%、8.9%、4.9%。‘湘苎3号’最佳生根培养基为附加TDZ (0.02 mg/L)+NAA(0.05 mg/L)+IAA (0.02 mg/L)的MS培养基,‘湘苎3号’、‘咸丰大叶绿’、‘A’...     

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.     

光敏核不育水稻（31301s）原生质体培养再生成株

从光敏核不育水稻３１３０１ｓ胚性细胞悬浮系分离培养原生质体，成功地实现了植株再生，原生质体在ＫＰＲ培养基中植板率达２．４％，在ＳＫＰＲ（除去ＫＰＲ中复杂的有机成分，以及葡萄糖以外的糖类）中获得了较高的植板率（１．８％），在高压灭菌取代过滤除菌的ＳＫＰＲ中也能培养成功。３１３０１ｓ原生质体再生细胞团直接分化的分化率达１０％，再生细胞团在含３％蔗糖和５００ｍｇ／Ｌ脯氨酸的Ｎ６培养基上增殖后再分化，可明     

Regeneration potential of CIMMYT durum wheat and triticale varieties from immature embryos

Twenty‐five durum wheat elite advanced lines and released varieties, and five triticale varieties were evaluated for their ability to produce embryogenic callus using three different media. For callus initiation and maintenance there were basal Murashige and Skoog (MS) medium containing double strains of macroelements and 2.5 mg/l 2,4D (DW1), basal MS medium containing 2.0 mg/l 2,4D (DW2), or basal MS medium supplemented with 1.0 mg/l 2,4 D and coconut milk (DW3). Plant regeneration was achieved on basal MS medium with indoleacetic acid and 6‐benzylaminopurine, and plants rooted on MS with 1‐naphthale‐neacetic acid. DW3 medium proved better than the other media tested for embryogenic callus initiation and maintenance. Regeneration rates varied widely with both genotype and initiation medium, with values ranging from no regeneration to 100% regeneration; the plantlets produced per embryo ranged from five to 20. Fourteen of the durum wheat genotypes showed 63–100% regeneration from DW3 callus formation medium, four lines from DW1 medium, and two lines from DW2. Four of the triticale varieties had regeneration of 48–100% from DW3 medium. After six subcultures, over a 6‐month period, genotypes lost their ability to regenerate plants. Only 10 lines retained some plant regeneration potential but regeneration was at reduced levels. Successful regeneration of durum wheat and triticale varieties will be used as an integral part of the transformation process.     

Callusing and Regeneration of Plantlets via Somatic Embryogenesis from Inflorescence Cultures of Triticum aestivum L.: Role of Genotype and Long-Term Retention of Morphogenic Potential

In the present investigation we have succeeded in obtaining a high frequency of regeneration of plantlets via somatic embryogenesis from callus derived from immature inflorescence explains of Triticum aestivum var, ‘Sonalika’. The explants were cultured on MS medium supplemented with 2,4-D, casein hydrolysate and coconut milk. A large number of embryoids germinated, to form plantlets on the medium when 2, 4-D was omitted altogether or provided at low concentration. Plantlets were transferred to soil under natural environmental conditions and were shown to have the normal chromosome number of 2n = 6x = 42. Experiments with nineteen other varieties show that there is a marked effect of genotype both on initiation of callusing as well as on regeneration. So far ‘Sonahka’ has proved to be the most responsive among varieties tested by us. With callus of the variety ‘Sonalika.’, we also conducted an investigation on long-term retention of regenerative potential. During Song-term culture, for about 12 months, the morphogenic potential gradually diminished and was finally lost, but the regeneration potential could be restored by subculturing at very short intervals.     

First in vivo assessment of feeding value of Festulolium hybrids derived from Festuca arundinacea var. glaucescens and selection for palatability

This paper reports on the first results of feeding value experiments, assessed with sheep, of hybrids between tetraploid Italian ryegrass (Lolium multiflorum, 2n = 4×= 28) and a progenitor of tall fescue, Festuca arundinacea var. glaucescens (2n = 4×= 28). Three hybrid populations resulting from either high, low or no selection for palatability were compared with a variety of Italian ryegrass as control and two varieties of the cultivated hexaploid (2n = 6×= 42) tall fescue, one of which had been selected for high palatability. On average, the hybrid populations were found to be as palatable as the improved tall fescue with voluntary intake (VI), in vivo digestibility of organic matter (DOM), and net energy expressed in fodder units for milk, similar to the best palatable tall fescue and to Italian ryegrass. However, selection for improved palatability among hybrids did not result in any further improvement of other traits, except VI in the hybrid selected for high palatability which significantly exceeded VI of Italian ryegrass. In conclusion, intergeneric hybridization between L. multiflorum and F. arundinacea var. glaucescens led to an immediate increase of feeding value compared with selection for palatability in tall fescue. However, to improve feeding value of hybrids further, similar selection for high palatability appeared questionable and possibly detrimental for other agronomic traits because of complicated inheritance in tetraploid L. multiflorum×F. glaucescens hybrids.     

Influence of genotype and culture medium on callus formation and plant regeneration from immature embryos of Triticum turgidum Desf. cultivars

Twelve durum wheat cultivars were evaluated for their response to in vitro tissue culture. Zygotic immature embryos were used to induce callus formation using four different Murashige and Skoog‐based media. Each contained 9.05 μM 2,4‐dichlorophenoxy acetic acid but differed in their carbon source (sucrose or maltose) and the presence of NaCl (0 mM or 40 mM). The influence of both genotype and medium on the type and percentage of callus produced was observed. Calli were either compact and frequently embryogenic, or soft and watery. Percentages ranged from 54 to 100%, depending upon genotype and induction medium. All calli were then plated on a regeneration medium containing 20 g/l sucrose, 2.68 μM 1‐naphthaleneacetic acid and 2.22 μ 6‐benzylaminopurine. The regeneration of plantlets was higher from compact than from soft calli, with a strong dependence on genotype and type of induction medium used. MSm induction medium (30 g/l maltose) and MS40s (30 g/l sucrose plus 40 mM NaCl) were best for inducing compact calli, and gave the highest proportion of regenerated plants. The in vitro response (number of total shoots from a compact callus/number of embryos plated) was higher for immature embryos of ‘Baztan’, ‘Bradano’ and ‘Don Pedro’. These cultivars are a good starting material for experiments involving transformation of calli from zygotic immature embryos.     

裸燕麦悬浮细胞系的建立及植株再生

对裸燕玫成熟胚进行离体培养。研究表明,基因型和不同激素组合,地成熟胚愈伤组织诱导具有重要作用。晋燕８号愈伤组织诱导率高达８５．７％,２,４－Ｄ是诱导愈伤组织必不可少的因素。采用循环培养法对愈伤组织进行改造,获得了易分散,颗粒状胚性愈伤组织,缓减了愈伤组织生活力衰退的速度。将松脆愈伤组织置于流体培养基中进行悬浮增减得到分散性。生长快的悬浮细胞系,悬浮系经分化培养获得的再生植株     

植物原生质体再生细胞壁研究进展

原生质体作为单细胞且具有全能性,为研究植物细胞壁生物合成提供了独特的视角和模型.原生质体细胞壁再生过程复杂,涉及诸多细胞信号转导通路,应用普通的研究方法难以全面解析整个再生壁过程的分子网络.为充分了解植物原生质体再生壁内在特性,推动植物原生质体再生壁的分子机理进一步完善,笔者从原生质体再生细胞壁培养、细胞壁再生的细胞生...     

新陆早32号、33号的体细胞胚胎发生和植株再生比较研究

以新疆主栽品种新陆早32号、33号和对照YZ1为研究材料,通过不同浓度的激素组合成功诱导获得了体细胞胚,并进一步发育成苗.研究发现,所用的12种激素组合均能有效诱导愈伤组织.虽然IBA+KT组合有利于诱导YZ1和新陆早33号的快速分化,但在增殖生长过程中体细胞胚容易褐化、死亡:在2,4-D+KT组合中,0.1 mg·L...