培养基及植物激素对烟草原生质体再生植株的影响

本文研究了不同基本培养基、培养方法及培养基中不同激素浓度配比对烟草叶肉细胞细胞原生质体再生植株的影响.结果表明,利用MS基本培养基进行琼脂糖固体平板培养,对烟草叶肉原生质体的发育速度、分裂频率及愈伤组织形成效果最佳;培养基(1/2 MS NAA 0.5 mg/L 2,4-D 1.0mg/L 6-BA0.5 mg/L 1.2%LMA) (1/2MS 2,4-D2.5mg/L KT0.5mg/L)的培养效果最好,再生细胞团的数目及愈伤组织最多;用1/2 MS IAA 2.0 mg/L KT 0.5 mg/L和1/2 MS IAA0.5 mg/L KT 2 mg/L两种培养基对愈伤组织诱导分化时出芽率高;在再生芽诱导生根时直接用不含任何激素的MS基本培养基效果最理想.     

水稻原生质体愈伤组织再生植株培养程序的比较

在4种不同的培养程序中应用了几种处理方法, 并对其在诱导水稻原生质体起源的愈伤组织 再生植株中的效果进行了比较。 直接将愈伤组织从含有2,4-D的增殖培养基转移到含有BA 和NAA的植株再生培养基上培养, 只能得到少量的弱苗(第1种程序)。 在增殖培养基中添加 ABA诱导了结节状的愈伤组织形成, 使愈伤组织的植株再生能力明     

巴戟天胚乳诱导愈伤组织及三倍体再生

目的:利用巴戟天种子内胚乳为外植体进行离体培养获得三倍体。方法:选取未成熟巴戟天种子内胚乳,置于不同培养基上诱导愈伤组织和分化再生植株,并对植株倍性进行根尖染色体鉴定。结果:在MS+2,4-D(2,4-二氯苯氧乙酸)2.0 mg/L+6-BA(6-苄基嘌呤)1.0 mg/L培养基上诱导获得愈伤组织,诱导频率达58.9%。愈伤组织在MS+6-BA1.0~2.0 mg/L+IBA 0.1~0.5 mg/L分化培养基上分化出不定芽。不定芽在1/2 MS+IBA 0.5 mg/L上诱导生根,生根率100%。再生植株根尖细胞染色体数目为2 n=3 x=33。结论:利用巴戟天胚乳培养获得三倍体是创造多倍体新种质的有效途径。     

香茶藨子茎尖愈伤组织诱导及植株再生

试验以香茶藨子茎尖为外植体,建立了愈伤组织诱导及植株再生体系。结果表明,诱导愈伤组织的最佳培养基为MS+NAA 0.5 mg/L+2,4-D 0.8 mg/L+6-BA 1.0 mg/L,诱导率为87.0%,愈伤呈紧密块状,生长良好;愈伤组织分化最佳培养基为MS+KT 1.5 mg/L+IBA 0.3 mg/L,愈伤分化率为55.53%;以1/2MS基本培养基添加IBA 0.5 mg/L,最利于生根培养。该体系的建立为香茶藨子规模化生产及遗传转化提供了技术平台。     

章丘大葱子叶再生体系建立及再生株倍性鉴定

以章丘大葱子叶为材料,研究了不同激素配比对愈伤组织诱导和继代、芽分化、芽点的伸长及根诱导的影响,并对获得的再生植株进行了鉴定。结果表明,诱导章丘大葱子叶产生愈伤组织、愈伤组织的继代培养、愈伤组织芽分化、芽点的伸长及诱导生根的最佳培养基及激素配比分别为：MS+2,4-D 2.0mg/L+KT 0.5mg/L、MS＋2,4-D 2.5mg/L＋KT 0.5mg/L、MS+2,4-D 0+KT 1.0mg/L、MS+KT 0.5mg/L和MS基本培养基;试管苗经驯化移栽后,共有22株成活,成活率为92%,对成活的再生植株进行形态学、叶片气孔及根尖染色体鉴定表明,共有3株发生变异,其中2株为外部形态上的变异,而另一株为染色体的加倍变异。     

大叶黄杨幼茎愈伤组织诱导的研究初报

以大叶黄杨的幼茎段为外植体,在添加BA和NAA、IBA、2,4-D不同激素组合的MS培养基上培养,对其愈伤组织进行诱导试验。结果表明：①生长素对愈伤组织诱导的效应为2,4-D >NAA>IBA;②在不同激素组合培养基上均可诱导出愈伤组织,其中MS+BA1.0~2.0 mg/L+NAA 1.0~1.5 mg/L、MS+BA1.5~2.0 mg/L+IBA1.0~1.5 mg/L、MS+BA0.5~1.0 mg/L+2,4-D 0.5~1.5 mg/L等对愈伤组织的诱导效果最好,其诱导率分别为74.3%、65.3%、81.1%。因此,通过科学配制不同激素组合的MS培养基,就能有效地诱导出大叶黄杨幼茎的愈伤组织。     

甘薯（Ipomoea batatas（L.）Lam.）及其近缘野生种原生质体的…

对甘薯品种高系１４号及其近缘野生种Ｉ．ｔｒｉｌｏｂａＬ．和Ｉ．ｌａｃｕｎｏｓａＬ．进行原生质体植株再生研究。从离体培养植株的叶柄分离出原生质体，将其培养在含有０．００５ｍｇ／Ｌ２，４－Ｄ和０．５ｍｇ／Ｌ激动素（ＫＴ）的ＭＳ培养基中，从原生质体获得了高频率的愈伤组织。培养８－１２周后，将直径达２－３ｍｍ的小愈伤组织转移到添加０．０５ｍｇ／Ｌ２，４－Ｄ的ＭＳ培养基上，转移３－６周后，将愈伤组织进一步转     

珍稀濒危植物银鹊树胚性细胞悬浮系的建立和植株再生

以银鹊树未成熟种子为试材,对银鹊树胚性愈伤组织诱导和胚性细胞悬浮培养的最佳培养条件进行研究,初步建立了银鹊树胚性细胞悬浮系与植株再生体系。将银鹊树未成熟种子子叶胚接种在添加1.0 mg/L 2,4-D、0.5 g/L活性炭的MS基本培养基上,诱导胚性愈伤组织。将诱导得到的胚性愈伤组织置于添加0.2 mg/L 6-BA、0.05 mg/L NAA和3 g/L蔗糖的MS液体培养基上振荡培养,由此建立了增殖速度快、分散程度好、稳定性较强的胚性细胞悬浮体系。将悬浮培养获得的子叶胚转到不含任何植物生长调节物质的MS固体培养基中,可以长成正常植株。     

降香黄檀愈伤组织培养与植株再生研究

为了实现降香黄檀工厂化快速繁殖和扩大栽培,并为降香黄檀抗寒基因导入打下一定的基础,以降香黄檀无菌实生苗茎段、叶片、根尖为外植体,MS为基本培养基,对各器官愈伤组织诱导与分化的最适培养基成分进行了研究。结果表明,可从降香黄檀无菌实生苗茎段、叶片、根尖成功地进行愈伤组织培养和植株再生。茎段、叶片、根尖诱导愈伤组织的最适培养基分别为1/2MS+6-BA 1.5 mg/L+NAA 0.10 mg/L、1/2MS+6-BA 0.5 mg/L+NAA 0.10 mg/L和1/4MS+6-BA 1.5 mg/L+NAA 0.05 mg/L;茎段、叶片、根尖的愈伤组织诱导丛生芽最适培养基分别为1/2MS+6-BA 1.5 mg/L+2,4-D 4.0 mg/L、1/2MS+6-BA 1.5 mg/L+2,4-D 3.5 mg/L和1/2MS+6-BA 1.5 mg/L+2,4-D 3.5 mg/L;茎段、叶片、根尖来源的单芽,其最佳生根培养基分别为MS+NAA 1.5 mg/L、MS+NAA 1.0 mg/L和MS+NAA 2.0 mg/L。比较茎段、叶片与根尖的愈伤组织诱导率、丛生芽诱导率和单芽生根率,得出以无菌实生苗根尖作为外植体是降香黄檀愈伤组织培养和植株再生的最佳选择。     

魔芋愈伤液体培养与植株再生的研究

以魔芋颗粒状愈伤组织为试材,进行了液体培养及植株再生诱导的探讨。结果表明,MS BA 1.0 mg/L 2,4-D 0.2 mg/L 蔗糖30 g/L 琼脂6.0 g/L培养基适合从芽鞘诱导颗粒状愈伤组织。获得的愈伤组织切割成0.3cm左右在MS BA 0.05 mg/L NAA 0.1 mg/L 蔗糖30 g/L PVP 1.0 g/L液体培养基中振荡培养,建立液体培养体系。液体培养材料的生长曲线呈上升的半抛物线型,从第6天到第12天期间增重占整个培养周期增重的80%以上,随着液体培养材料的生长,培养液pH值上升。将液体培养12 d的材料转到1/2MS BA 0.6 mg/L NAA 0.1 mg/L 葡萄糖30 g/L 维生素C100 mg/L 琼脂6.0 g/L培养基上,进行愈伤组织的分化培养;然后将长到出叶期的大芽切割转入到1/2MS NAA 0.1 mg/L 蔗糖30 g/L 琼脂6.0 g/L生根培养基诱导生根,形成再生植株。     

甘薯和Ipomoea lacunosa的种间体细胞杂种植株再生及鉴定

用ＰＥＧ融合法融合甘薯品种高系１４号和近缘野生种Ｉｐｏｍｏｅａ ｌａｃｕｎｏｓａ的原生质体，将融合原生质体培养在含有０．０５ｍｇ／Ｌ２，４－Ｄ和０．５ｍｇ／Ｌ ＫＴ的ＭＳ培养基上，愈伤组织迅速增殖。将其中的７０个愈伤组织培养在添加３．０ｍｇ／Ｌ ＢＡＰ的ＭＳ培养基上，并进一步培养在ＭＳ基本培养基上，获得９株再生植株。过氧化物酶同工酶、酯酶同工酶和ＲＡＰＤ分析表明，其中２株再生植株（ＫＬ１和ＫＬ３）     

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     

花生原生质体分离与培养

以花生品种花育20为材料,探讨不同的酶液浓度、酶解时间及渗透压对花生原生质体分离的影响。结果表明:原生质体分离的适宜酶液配比是2%纤维素酶（Cellulase Onozuka RS）、0.2%果胶酶(Pectolyase Y-23),甘露醇渗透压调节剂的浓度为0.7 mol/L,暗处理10 h,叶片原生质体产量为4.86×105/ml,存活率75.6％,愈伤组织原生质体产量为4.97×105/ml,存活率75.4％。将分离的原生质体培养在添加1mg/LNAA和4mg/LBAP的改良MS液体培养基中。培养约2～3d后,细胞开始分裂。然后部分细胞继续分裂,并形成细胞团。培养5～6周后,将培养物转移到添加2mg/L2,4-D和3mg/LBAP的MS液体培养基（pH 5.8）中进行培养。7～8周后,将形成的直径为1～2mm的小愈伤组织转移到添加1mg/LNAA和5mg/LBAP的MS固体培养基上培养,小愈伤组织迅速生长。转移3～4周后,愈伤组织长至7～9mm。     

Plant regeneration from protoplasts of Iris germanica L.

Summary Protoplasts were isolated enzymatically from suspension cultures derived from embryogenic calli induced by leaf base culture of Iris germanica. In protoplast culture, the effects of glucose concentration, different sugars and combinations of 2,4-D and KIN on protoplast division and colony formation were examined. N6 medium supplemented with 0.1–1 mg/l 2,4-D, 1 mg/l KIN, 200mg/l casein hydrolysate, 250 mg/l proline, 0.2 M glucose and 20 g/l agarose was suitable for protoplast division and colony formation. When colonies formed were transferred onto hormone-free MS medium, many plantlets were regenerated through somatic embryogenesis. Thus, we could establish a plant regeneration system from protoplasts of I. germanica.Contribution from the Laboratory of Plant Breeding, Faculty of Agriculture, Miyazaki University, Japan, No. 95.     

Continuous Plant Regeneration from Established Embryogenic Cell Suspension Cultures of Italian Ryegrass and Tall Fescue

The suitability of different protocols was compared for entire plant regeneration by somatic embryogenesis, of the forage plants Lolium multiflorum Lam. (Italian ryegrass) and Festuca arundinacea Schreb. (tall fescue). In the first protocol, miniature embryos were used as starting material, while mature seeds were retained in the other two. Whichever the considered protocol, undifferentiated calli were produced on Murashige and Skoog MS medium supplemented with 2,4-D. The calli were subcultured in the dark on solid MS agar medium, containing 5 mg/1 2,4-D (protocol 2) or on solid MS medium followed by transfer to a rotated liquid MS medium with 2 mg/1 2,4-D (protocol 1). In these conditions, induction of somatic embryogenesis occurred, and whole plants were regenerated during a limited lapse of time, upon transfer in the light, to MS medium supplemented with BAP but devoid of 2,4-D. The simultaneous elimination of 2,4-D and transfer to light appeared essential for full regeneration of the plants. Using this characteristic, an additional step was added to a new protocol (protocol 3) in which microcalli, cultured on liquid MS medium containing 5 mg/1 2,4-D, were transferred to the same medium with 2 mg/1 2,4-D, in the dark. In these conditions, the suspensions kept their embryogenic potential for months. In all cases, plantlets were successfully transferred into the soil. An evaluation of the somaclonal variation potential of the plants issued from each protocol is now underway.     

大豆幼荚子叶原生质体培养及植株再生

本文研究了13个栽培大豆（Glycine max L.）品种原生质体培养的再生能力。从大豆幼荚子叶酶解游离原生质体,用Gellan Gum进行株状包埋,悬浮在含2,4-D 0.1-0.2mg/L,BA0.5-1.0mg/L的改良MS液体培养基中,原生质体培养3天后开始第一次分裂,以后持续分裂。供试基因型间的10天植板率差异显著,变幅为33-67%。30天内形成大量的     

Plant regeneration from embryo-callus culture in barley

Summary Plants were regenerated from callus cultures initiated from immature embryos of barley, Hordeum vulgare L. Immature embryos from seven diverse genotypes were cultured on modified Murashige and Skoog (MS) medium supplemented with 1.5 mg 2,4-D and 6.5 mg IAA/l. Of the 249 embryos cultured, 30% initiated callus within 8 days. Subculture of callus for 80 to 100 days on half-MS medium supplemented with 0.5 mg/l 2,4-D and 1.0 mg/l zeatin resulted in organogenesis. Culture of organogenic calli for 30 days on half-MS medium without growth regulators produced plants which originated mostly via multiple shoot formation. Callusing response of the tested genotypes ranged from zero to 44%; however, only 23% of the calli were regenerative. Regenerated plants included variants for chlorophyll deficiency, plant height, stem thickness, spike shape, pollen fertility, seed set and ploidy.     

新疆大叶紫花苜蓿(Medicago sativa.L)子叶、下胚轴、根的植株再生

随着植物抗逆性研究和植物转基因技术的发展，通过异源目的基因转化培育耐盐碱苜蓿品种的研究已引起人们的关注，植物受体高频再生体系的建立是异源转化高效的基础。选取新疆大叶紫花苜蓿种子萌发5~7d无菌苗的子叶、下胚轴及根为外植体，诱导愈伤培养基为MS+2,4-D 0.1~3.0 mg/L(8种不同水平)或MS+2,4-D 2.0 mg/L+ KT 0.01~0.5 mg/L(10种不同水平)，诱导芽培养基为MS+6-BA 0.5 mg/L+ NAA 0.05 mg/L，生根培养基为MS。结果表明，外植体在MS+2,4-D 2.0 mg/L+ KT 0.2 mg/L培养基中能够产生状态较好可再分化的愈伤组织，子叶、下胚轴、根的平均出愈率分别为93.1%、100%、100%。愈伤组织在MS+6-BA 0.5 mg/L+ NAA 0.05 mg/L培养基中培养40~80d中均可分化芽，子叶、下胚轴、根的芽平均分化率为50%、78%、50%，将2 cm以上的芽转入MS培养基中诱导生根，14d后，生根的小植株炼苗移入花土中，成活率达90%以上。子叶、下胚轴、根在该体系中均能获得再生植株，根也是一种较好的植株再生材料，以根为外植体进行植株再生的研究报道还较少。     

Effects of growth regulators on callus cell growth,plant regeneration,and somaclonal variation of smooth bromegrass (Bromus inermis Leyss.)

Summary Immature inflorescences of smooth bromegrass were cultured on MS agar media supplemented with varying combinations of 2,4-D and kinetin. Callus was initiated from segments of young inflorescences on each medium. All of the calli were subcultured monthly for 5–6 times and transferred onto hormone-free MS medium for plant regeneration. Addition of kinetin to the basal medium stimulated shoot initiation in the callus cultures. Plantlets were regenerated only from calli grown on media containing 2 and 6 mg I^-1 2,4-D with a supplement of 0.2 mg I^-1 kinetin. No albino plantlets were produced. Morphological characteristics and dry matter yield of ten somaclones and the parental plant (SBG7) were studied in the greenhouse in a randomized complete block experiment with five replications. There was significant variation (P>0.01) among genotypes for all morphological characteristics studied. Although all somaclone heights and leaf widths were lower than those of the parental plant, the somaclone F9A, F10A, and F10B had larger tiller numbers, and leaf/stem ratio by dry weight than the parental plant. Only somaclone F9B gave higher specific leaf area and leaf area ratio than the parental plant. Almost all somaclones had the same leaf length, total dry weight, and specific leaf weight as the parental plant. The variation found in somaclones should permit selection for desirable agronomic traits.     

Callus induction and plant regeneration from immature and mature embryos of winter durum wheat genotypes

Seven genotypes of winter durum wheat (Triticum durum Desf.) were cultured to establish an efficient method of callus formation and plant regeneration from mature embryo culture, and to compare the responses of immature and mature embryo cultures. Immature embryos were aseptically dissected from seeds and placed, with the scutellum upwards, in dishes containing Murashige and Skoog's (MS) mineral salts and 2mg 2,4- dichlorophenoxyacetic acid (2,4-D) per litre. Calli and regenerated plants were maintained on 2,4-D-free medium. Mature embryos were moved slightly on the imbibed seeds. For callus formation, the seeds with moved embryos were placed, furrow downwards, in dishes containing 8 mg 2,4-D per litre. The developed calli and regenerated plants were maintained on the MS medium. Plants regenerated from both embryo cultures were vernalized and grown to maturity in soil. Variability was observed among the wheat genotypes tested for various culture responses in both explant cultures. Callus induction rate and regeneration capacity of callus were independent of each other. Mature embryos have a low frequency of callus induction but a high regeneration capacity. Considering availability, rapidity and reliability, this form of mature embryo culture can be used as an alternative method for immature embryo culture.