不同激素对花生离体分化的影响

对TDZ和2,4-D等激素在花生成熟胚外植体分化中的影响进行了研究.结果表明,花生成熟胚3～5 d龄实生苗的幼叶和胚轴在低浓度TDZ的诱导下,可分化产生高频不定芽和少量体细胞胚,转到无激素MS培养基或MS BA 0.5 mg/L NAA 0.4 mg/L的培养基后形成丛生苗.丛生苗分离后转入含1/2 MS(大量元素) IBA 0.4 mg/L的培养基中诱导生根,可形成完整的再生植株.幼叶分化率高于胚轴,但胚轴分化成苗速度快.无菌水浸泡16～24 h的胚轴在5～ 30 mg/L 2,4-D的诱导下,分化产生低频不定芽;而胚叶则产生高频体细胞胚,但畸形较严重.     

花生胚小叶离体再生体系的优化研究

目的：研究培养基中植物激素的种类、浓度和基因型差异对花生胚小叶离体培养再生效率的影响。方法：以花生胚小叶为外值体,比较各处理间的芽丛诱导率、芽丛出苗率和植株再生率,确定每个品种对应的适宜培养基。结果：小花生品种丰花2号的适宜芽丛诱导培养基为MSB＋3.0mg/LBA＋0.7mg/LNAA,芽丛诱导率为81.49%,再生率为94.39%;适宜芽丛成苗培养基为MSB＋2.5mg/LBA,芽丛出苗率为172.00%,再生率为122.10%。大花生品种丰花3号的适宜芽丛诱导培养基为MSB＋4.5mg/LBA＋0.2mg/LNAA,芽丛诱导率为72.33%,再生率为10.70%;适宜芽丛成苗培养基为MSB＋5.0mg/LKIN,芽丛出苗率为33.33%,再生率为4.03%。结论：不同基因型之间的成苗芽丛百分率、芽丛出苗率和植株再生率差异显著。     

‘SK4—316’胡萝卜体胚的诱导和培养

以'SK4-316'胡萝卜无菌苗的下胚轴为外植体,研究不同培养基配方和培养条件对愈伤组织诱导、体细胞胚间接发生及其同步化培养的影响,以及不同脱分化时间、脱分化培养基及外植体续存时间对体细胞胚直接发生的诱导及其培养的影响.结果表明:含3%蔗糖、0.8%琼脂的1/2MS + 2,4-D 2.5 mg/L + 6-BA(或KT)0.5 mg/L + CH 300 mg/L是诱导愈伤组织的良好培养基;1/2MS + 2,4-D 1.25 mg/L + KT 0.25 mg/L + 6-BA 0.25 mg/L(含3%蔗糖)适于愈伤组织分化并诱导体胚发生,0.02% ABA对体胚的诱导有促进作用,0.06% ABA或15% PEG能促进体胚成熟;外植体在MS + 2,4-D 1.0 mg/L固体培养基上脱分化培养48 h,再转入MS + CH 300 g/L液体培养基中可诱导体胚直接发生,但随着外植体续存于诱导培养基中时间的延长,体胚发生变异的几率也渐增.     

2,4-D诱导的花生体细胞胚发生的组织学研究

花生成熟胚胚叶在佃附加20mg/L 2,4-D的培养基上诱导20d后,转移至无激素培养基MS0继续培养,可获高频体细胞胚发生。组织学观察表明,体细胞胚起源于胚叶上表皮及表皮下数层细胞,这些细胞脱分化形成细胞质浓厚、细胞核大的胚性细胞团,胚性细胞团继续分裂形成体细胞胚。体细胞胚的发育过程经历球形胚、心形胚、鱼雷胚、子叶胚四个时期。     

小麦遗传转化受体系统建立的研究

选用‘小偃22’和‘宁春16’小麦品种的成熟胚和幼胚进行培养,研究不同种类的胚和培养因子对愈伤组织诱导和分化的影响。结果表明,幼胚和成熟胚的愈伤组织诱导率无明显差异,但较高浓度的2,4-D有利于成熟胚的诱导,而幼胚培养时2,4-D浓度的影响效果因品种而异;两种外植体分化率的高低与KT/IAA的配比均有密切关系,但高浓度的激素水平不利于成熟胚的分化;诱导培养基中低浓度的2,4-D有利于所诱导的愈伤组织的分化。同时,在诱导培养基中添加低浓度的KT能显著提高两品种成熟胚愈伤组织的分化率;各种培养基处理与品种间都存在显著的互作效应,‘小偃22’成熟胚培养的最佳培养基组合为MSD 3.0 mg/L 2,4-D和MSD 0.5 mg/LIAA 1.0 mg/L KT,幼胚培养为MSD 4.0 mg/L 2,4-D和MSD 0.5 mg/L IAA 1.0 mg/L KT;‘宁春16’成熟胚培养为MSD 4.0 mg/L 2,4-D和MSD 1.0 mg/L IAA 1.0 mg/L KT,幼胚培养时为MSD 1.0 mg/L 2,4-D和MSD 2.0 mg/L IAA 2.0 mg/L KT。     

小麦成熟胚愈伤组织诱导及分化研究

以2个小麦品种成熟胚为外植体进行离体培养,研究了不同预处理、不同2,4-D浓度及与KT组合、不同蔗糖浓度等因素对愈伤组织诱导及分化的影响。结果表明:4℃低温预处理可提高愈伤组织的出愈率及再生苗率,2个材料的出愈率及再生苗率均达到90%和30%以上;在不同预处理条件下,2,4-D浓度对出愈率及再生苗率的影响与基因型有关,2,4-D浓度为1~2 mg/L更有利于愈伤组织诱导及分化;附加KT能缓解高浓度2,4-D对再生苗率的抑制作用,而对于在1、2 mg/L 2,4-D的培养基中附加KT则不表现这种作用;蔗糖浓度则在30 g/L条件下更有利于愈伤组织诱导。因此通过4℃低温预处理,在MS基本培养基中附加1~2mg/L 2,4-D及30 g/L蔗糖亦可促进小麦成熟胚愈伤组织的诱导和分化。     

云南大叶茶体细胞胚发生及体细胞胚苗形成体系的建立

利用云南大叶茶(Camellia sinensis var.assamica Kitamura)胚性细胞系(CL_1)中悬浮培养物,建立了高频率同步化体细胞胚发生及体胚苗形成体系。以改良的MS为基本培养基,将CL_1中培养物由液体保持培养基(0.1mg/L 2,4-D 0.5mg/L 6-BA)继代转入液体诱导培养基(0.05mg/L 2,4-D 0.50mg/L6-BA),暗培养诱导28d,转入不含任何激素的液体分化培养基中再培养28d,获得了不同发育时期的体细胞胚,其发生频率为81.5％。不同发育时期的体细胞胚用不同目的细胞筛收集,在液体生长培养基(1/2 MS 1.0mg/L GA_3 0.5mg/L 6-BA)中培养发育成熟。ABA有利于高质量体细胞胚的形成。20～70月大小的体细胞胚在固体生长培养基中成苗转换率为75％。在液体悬浮培养条件下观察记录了体细胞胚发育过程,证实其过程与合子胚的形态发生过程相似。     

几种匍匐翦股颖成熟胚愈伤组织诱导及植株再生

利用成熟种子作为外植体,分析了2,4-D对匍匐翦股颖胚性愈伤组织诱导与植株再生体系的影响,并对体细胞胚的发生过程进行了观察.实验结果表明,在2.0 mg/L 2,4-D 0.1 mg/L 6-BA时胚性愈伤组织的诱导频率最高.随着2,4-D浓度的增加,愈伤组织的诱导和分化能力明显下降.在再生过程中采用1.0 mg/L的6-BA达到了比较好的效果,愈伤组织的再生频率大部分在90%以上.同时发现适当提高肌醇浓度可以使苗长得较为粗壮.在实验中发现匍匐翦股颖的体细胞胚发生过程为球形胚、心形胚、鱼雷胚和子叶胚.     

防风组织培养中畸形胚状体的发生和控制

采用组织培养和石蜡切片法,分析多种因素对防风畸形胚状体发生和发育过程的影响及控制。结果表明,诱导正常体细胞胚高频发生的培养基组合是：启动胚性愈伤组织的培养基是MS 0．5mg／L 2．4-D,蔗糖浓度3％;分化培养基是MS 1mg／L 6-BA 0．2mg／L NAA 0．5mg／L ABA,蔗糖浓度4％～5％;成熟培养基是MS培养基,蔗糖浓度3％。结论：一定浓度的生长素是诱导防风胚性愈伤组织的关键因素,细胞分裂素在体细胞胚的分化和发育过程中起协同作用;蔗糖浓度、ABA、接种方法以及适宜的培养条件和培养容器等均可有效降低畸形胚状体的发生率。     

ABA诱导毛酸浆下胚轴体细胞胚状结构的直接发生

研究了ABA与2,4-D等植物激素对毛酸浆组织培养的影响。其中,以浓度为1.0mg/L的2,4-D对毛酸浆下胚轴愈伤组织诱导结果最好。在子叶、子叶柄、下胚轴以及胚根四者当中,下胚轴对2,4-D的反应状况最好。结合ABA与2,4-D在MS培养基上能够有效地诱导出高质量的球形胚状结构。经过4周的培养,在2.0mg/L2,4-D和1.0mg/L ABA组合的情况下,球形胚状结构的诱导频率可达96.7％,每块外植体平均能产生6.7个球形胚状结构。球形胚状结构可以至少在5代以内反复发生。进一步讨论了ABA对毛酸浆体细胞胚发生的影响以及体细胞胚发生与环境胁迫的可能关系。     

苗期灌水量对花生生理特性和产量的影响

在人工控水条件下,研究了苗期不同灌水量对两个抗旱性不同的花生品种“农大818”和“鲁花11”生理特性及产量的影响.结果表明：随着灌水量的减少,花生叶片光合速率逐渐下降,叶片丙二醛(MDA)含量增加;灌水60～80 mm（适当干旱）可增加叶片中超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性,提高叶片可溶性蛋白质(P_r)含量;干旱解除后,叶片中SOD、POD、CAT活性和P_r、MDA含量显著降低,而光合速率显著升高.随着灌水量的减少,花生荚果和籽仁产量降低,但农大818的降低幅度不如鲁花11明显,表明农大818比鲁花11更耐旱;在节水条件下,鲁花11苗期适宜灌水量不能低于80 mm,农大818不能低于60 mm.     

Plant regeneration from protoplasts of soybean (Glycine max L.)

Protoplasts were isolated from immature cotyledons of six cultivars of Glycine max L. and cultured in the KP8 liquid medium supplemented with 0.2 mg/L 2,4-D, 1 mg/L NAA and 0.5 mg/L ZT. The protoplasts started to divide after 3–5 days of culture. Sustained divisions resulted in mass production of cell colonies and small calli in 6 weeks. The calli further grew to 2–3 mm on the gelritesolidified K8 medium and were transferred onto the MSB medium with 1 mg/L 2,4-D and 0.25 mg/L BA, to obtain compact and nodular calli. Shoot formation was initiated on MSB medium with 0.15 mg/L NAA, and BA, KT and ZT, 0.5 mg/L of each, with or without 500 mg/L CH. It was followed by plant regeneration. So far, 87 plants have been regenerated from 4 cultivars, and normal seeds were obtained from them after transplanting into pots.Abbreviation IAA indol-3-acetic acid - NAA naphthalene acetic acid - 2,4-D 2,4-dichlorophenoxy acetic acid - KT kinetin - BA 6-benzyladenine - ZT zeatin - CH casein hydrolysate     

High efficiency <Emphasis Type="Italic">in vitro</Emphasis> plant regeneration from epicotyl explants of Chinese peanut cultivars

An efficient organogenesis and micropropagation system was developed for in vitro plant regeneration of multiple cultivars of peanut (Arachis hypogaea). The system was used to regenerate plants from nine cultivars: Luhua no. 9, Luhua no. 13, Luhua no. 14, Fenghua no. 1, Fenghua no. 3, Fenghua no. 5, Huayu no. 23, Qinglan no. 2, and Baisha 1016. Epicotyl and embryo axis explants were cultured on Murashige and Skoog (MS) basal medium supplemented with 33.29–44.39 μM 6-benzyladenine (BAP) and 2.15–4.30 μM α-naphthaleneacetic acid (NAA). The highest rate of shoot formation was observed in epicotyl explants taken from 4-d-old seedlings (5.1 ± 1.4 shoots per explant). Optimum shoot development was observed in explants cultured on MS medium containing 4.48 μM BAP and 2.89 or 5.78 μM gibberellin (GA₃). Well-developed shoots (3–5 cm high) formed roots after 2 wk on MS medium containing 0–2.69 μM NAA. We observed that all multiple shoots formed at the site of epicotyl incision and at the upper end of each section, indicating physiological polarity of shoot formation. The maximum shoot induction rate for Luhua no. 9, Luhua no. 13, Luhua no. 14, Fenghua no. 1, Fenghua no. 3, Fenghua no. 5, Huayu no. 23, Qinglan no. 2, and Baisha 1016 was 60.0%, 83.3%, 80.7%, 91.5%, 86.0%, 59.7%, 75.0%, 67.3%, and 72.7%, respectively. This regeneration system will play a vital role in achieving the genetic improvement of Chinese peanut.     

Effect of 2,4-Dichlorophenoxyacetic Acid and NaCl on the Establishment of Callus and Plant Regeneration in Durum and Bread Wheat

Optimal callus induction and plant regeneration were obtained in bread and durum wheat by manipulating the NaCl concentration in the induction medium. Immature embryos from a high regeneration line of spring wheat (Triticum aestivum L.), 'MPB-Bobwhite 26', and an elite durum wheat (Triticum turgidum var. durum L.), 'Mexicali', were cultured in E3 induction medium consisting of Murashige and Skoog (MS) medium, 2.5 mg l^–1 2,4-dichlorophenoxyacetic acid (2,4-D), 2% sucrose and 0.9% Bacto agar. The treated embryos were transferred to E3 liquid medium supplemented with various levels of 2,4-D and NaCl. Incubation on medium containing 2.5 mg l^–1 2,4-D for 45 days produced callus and plant regeneration in 'MPB-Bobwhite 26', but lower callus yield and plant regeneration in 'Mexicali', indicating that 2,4-D alone was not sufficient for callus induction and plant regeneration in this durum variety. Callus yield and regeneration frequencies were higher in 'Mexicali' embryos that were incubated in media containing 2 mg l^–1 2,4-D and 2 mg l^–1 NaCl. The presence of NaCl in the medium beyond the initiation phase was detrimental to plant regeneration. The use of NaCl in the callus formation could form the basis for improved transformation of durum wheat varieties.     

花生体细胞胚的诱导及其植株再生

采用不同成熟度的花生胚轴为外植体进行体细胞胚诱导及植株再生研究,结果表明,成熟胚轴在高浓度2,4－D的MS培养基中,经过30d左右的培养,可直接诱导产生出大量的体细胞胚,含40mgL~－12,4－D的培养基中体细胞胚的诱导率达100％,平均每个外植体产生11．58个体细胞胚．体细胞胚的继代培养需降低2,4－D的浓度（1－20mgL~－1）．未成熟胚轴的体细胞胚诱导及继代培养的2,4－D浓度宜为10mgL~－1．将诱导的体细胞胚转接到合5－10mgL~－1BA的MS培养基中,体细胞胚能够萌发再生成无根小植株,将其转接到生根培养基中可获得完整小植株．     

玉米幼胚高效再生系统的建立

建立了玉米幼胚高效再生系统.经研究发现,苏玉1号、农大3138、农大108的幼胚培养在含有2,4-D(2 5 mg/L)的IM培养基上后,大多数幼胚能愈伤化并增大,形成基部相连、上部分开的微芽结构;微芽结构在转移到BM培养基上后,形成小植株;进一步转移到RM培养基上,它们长根并形成完整植株.玉米幼胚高效再生植株与下列因素有关玉米基因型、幼胚大小、幼胚长芽至分化时间、6-BA、IBA、Gelrite.不同品种玉米再生能力有显著差异,幼胚大小在1～2mm之间再生能力强,幼胚长芽至分化时间4～6 d最好.激素6-BA浓度在0.5～0.6 mg/L之间有利于微芽形成小植株,IBA浓度在0.6～1 0 mg/L促进生根.Gelrite可代替琼脂粉用于玉米生根.     

长柄双花木愈伤组织诱导及植株再生

以长柄双花木当年生嫩梢上的叶柄、嫩茎、嫩叶为外植体,对影响长柄双花木愈伤组织诱导和继代、分化主要因素进行研究。结果表明:在培养基MS+NAA0.5mg/L+2,4-D2.0mg/L上,三种外植体均可诱导出愈伤组织,其中叶片愈伤组织诱导率最高。该培养基还可作为愈伤组织继代培养基,但继代培养周期不超过2周。愈伤组织接种在MS+BA2mg/L上分化不定芽,根的诱导在1/2MS+IBA0.5mg/L培养基上进行。     

The effect of 2,4-dichlorophenoxyacetic acid and donor plant environment on plant regeneration from immature inflorescence-derived callus of Lolium perenne L. and Lolium multiflorum L.

Callus cultures were obtained from immature inflorescences of perennial ryegrass (Lolium perenne) and Italian ryegrass (Lolium multiflorum). Inflorescence segments were cultured on Murashige and Skoog medium (MS) supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D). The response in culture with regard to compact callus induction, embryogenesis and plant regeneration was determined for different varieties. The in vitro response was compared for explants from field-grown plants and explants from greenhouse-grown plants. The effect of different 2,4-D concentrations on the in vitro response was also investigated in one L. perenne variety and one L. multiflorum variety. The percentage of explants that formed compact callus and embryogenic callus differed strongly with the cultivar. There was no consistent effect of the growth conditions of the donor plants or the 2,4-D concentration of the medium on this response. Green plants were regenerated from all the cultivars tested. Explants from field-grown plants showed a higher tendency to form albino shoots than explants from greenhouse-grown plants. In the L. perenne variety tested higher 2,4-D concentrations (up to 15 mg/l) resulted in a lower regeneration frequency of green shoots and a higher regeneration frequency of albino shoots (up to 12.5 mg/l). In the L. multiflorum variety tested the effect of 2,4-D on regeneration was less pronounced.     

Plants regenerated from mesophyll protoplasts of white mulberry

INTRODUCTIONProtoplastcultureis0neofthen1ostrapidlydevel0pingareasinp1anttissueculture,becauseofitsimportancei11plantgeneticmanipulation.However,sofar,thereareonlyafewforesttreespeciesinwhichplantregenerationfr0mprotoplastshaJsbeensuccessful,namelyLiriode…     

Protoplasts Culture and Plant Regeneration of Soybean (Glycine max L. and G. soja)

Protoplasts were isolated enzymatically from immature cotyledons of soybean. The protoplasts divided to form calli in the K8P liquid medium. The calli further grew to 2–3 mm on the solid K8 medium and were transferred onto the MSB medium (MS minerals+B5 organic components+0.5–1.0 mg/l 2,4-D+0.2–0.5 mg/l BA) to obtain compact and nodular calli. Shoot formation was initiated on M1 medium (MSB medium with 0.15 mg/1 NAA, and BA, KT and ZT, 0.5 mg/l of each, 500 mg/1 CH). Differentiation frequency was 13.6–24.2%. Plants have been regenerated from protoplasts of immature cotyledons in 2 cultivars, and normal pods were obtained from them.