The Comparison in Tissue Culture Ability of Mature Embryo in Different Cultivars of Rice

In order to study the regeneration technology of mature embryos in different rice varieties,nine japonica,nine indica and eleven hybrid rice varieties of two line or three line or superiority combinations were selected as explants to study the callus induction,differentiation and regeneration rates on different media.The higher callus induction （61.7-89.2%） was observed in japonica rice,when cytokinin was added at lower concentration （0.3 mg L-1 6-BA） in M8 basal medium,supplemented with 30 g L-1 sucrose,8 g L-1 agar and 2 mg L-1 2,4-D.Further,the addition of two cytokinins （2 mg L-1 6-BA,0.5 mg L-1 KT） and 1 mg L-1 NAA in the M8 basal supplemented medium resulted in 9.1-100% of the callus induction in indica rice.The percent callus induction in hybrid rice varieties was 40-86.3% when addition of 1 mg L-1 6-BA and 1 mg L-1 KT was added,and the cytokinins was required by the japonica and indica rice varieties in the M8 basal supplemented medium.It was observed that when the 0.5 mg L-1 2,4-D and 1 mg L-1 6-BA were added in japonica rice,and 0.2 mg L-1 2,4-D and 0.5 mg L-1 6-BA were added in indica and hybrid rice in the MS different media,the regeneration rates were 9.2-59.5%,3.6-87.5% and 17.2-43.2% for japonica,indica and hybrid rice,respectively.Thus,the regeneration technology with higher output is established in the mature embryos of similar rice varieties.

Study on Plant Regeneration of Wheat Mature Embryos Under Endosperm-Supported Culture

To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars （Triticum aestivum L. cv.） were cultured with or without endosperm to test their efficiency of callus induction and plant regeneration. When embryos were cultured together with endosperm （endosperm-supported culture, ES）, the percentage of callus induction was significantly lower than that when embryos were cultured in the absence of endosperm （non-endosperm-supported culture, NES）. This pattern was evident in most genotypes, regardless of whether 2 or 8 mg L^-1 2,4-D was added in the NES culture. However, in ES culture, more induced calli were differentiated into distinct green spots and they further developed into plantlets. Thus, more plants were regenerated in ES culture than in the NES treatment. Most of the eight tested genotypes showed a significant difference in callus induction rate and plantlet regeneration in both ES and NES cultures. In addition, the enzymatic activity of oxalate oxidase in the callus of ES culture condition was obviously higher than that in the callus of NES culture condition, suggesting that the activity of oxalate oxidase may be a parameter for selection of calli with potential for plantlet regeneration. These results indicate that wheat mature embryos are valuable explants for highly efficient callus induction and plant regeneration, if proper treatment and medium are used.

Dicamba and Sugar Effects on Callus Induction and Plant Regeneration from Mature Embryo Culture of Wheat

To establish a highly efficient plant regeneration system for wheat genetic transformation, the effects of three different concentrations of dicamba and two different sugar types on callus induction and plant regeneration from mature embryo cultures were evaluated. Callus induction and plant regeneration were obtained from mature embryos of two commercial cultivars Zhoumai 18 and Yumai 34 （Triticum aestivum L.） cultured on L3 basal medium. The results showed that the efficiency of mature embryo culture was significantly influenced by the genotypes, sugar types and dicamba concentrations. 4 mg L^-1 dicamba proved the best effective for inducing embryogenic callus and also gave the highest proportion of plants regenerated across the two cultivars. Substitution of maltose by sucrose significantly improved the plant regeneration efficiency in both cultivars. There was a significant interaction between genotype-by-sugar types, and sugar types-bydicamba concentrations. Overall, Zhoumai 18 gave the highest frequency of plant regeneration （82.65%） when dicamba concentration was 4.0 mg L^-1 and with sucrose in initial callus induction. These results will facilitate genetic transformation work with elite wheat.

A Preliminary Study on Immature Embryo Culture and Plant Regeneration of Lagerstroemia indica

[ Objective ] This study aimed to explore immature embryo culture of Lagerstroemia indica and investigate the appropriate conditions for growth and differentiation. [ Method] Immature embryos of L. indica were employed as the explants for germination induction to establish aseptic lines. Based on that, the effects of different hormone levels and culture conditions on immature embryo culture of L. indica were analyzed. [ Result ] Peeled immature embryos of L. indica were germinated easily, leading to a germination rate of 100%. The optimal initial medium was MS ＋ BA0.5 ＋ NAA0.1 ＋ sucrose 3.0% ＋ agar 0.7% ; the optimal shoot induction medium was MS ＋ BA0.5 ＋ NAA0.1 ＋ sucrose 3.0% ＋ agar 0.7% ＋ coconut milk 10% ; the optimal rooting medium was MS ＋ BA0.5 ＋ IBA0.1 ＋ sucrose 3.0% ＋ agar 0.7% ＋ coconut milk 10%. [ Conclusion] This study provided a technical reference for subsequent optimized breeding of L. indica.

Effects of CuSO_4 and Uniconazole on Mature Embryo Culture in Japonica Rice

In order to establish the system of high frequency plant regeneration for japonica rice mature embryos, the effects of different concentrations of CuSO4 and uniconazole on in vitro culture of mature embryos were studied using three rice cultivars of Kongyu 131, Longjing 24, and Dongnong 425 as test materials. The results showed that callus induction and differentiation of japonica rice mature embryos were apparently improved on the medium with 10-15 μmol·L-1 CuSO4 and 0.50-1.00 mg·L-1 uniconazole. Induction and differentiation rates of different genotype rice mature embryos displayed different sensitivities to CuSO4 and uniconazole. For the callus induction frequency of three varieties, the optimal concentration of CuSO4 was 15.0 mol·L-1. When the concentration of CuSO4 was 15 μmol·L-1, the plantlet differentiation rates of Kongyu 131 and Dongnong 425 got to the highest, while the concentration of CuSO4 was 10 μmol·L-1 for Longjing 24. For the callus induction and plantlet differentiation rates of Kongyu 131 and Dongnong 425, the ideal concentration of uniconazole was 0.50 mg·L-1 and for Longjing 24 was 1.00 mg·L-1.

Plantlets Regeneration via Somatic Embryogenesis from the Nucellus Tissues of Kinnow Mandarin (Citrus reticulata L.)

Studies were initiated to explore the role of nucellus tissues and growth regulators in plantlets regeneration via somatic embryogenesis of Kinnow mandarin [Citrus reticulata L. (Blanco)]. Nucellus tissues were cultured on MS medium supplemented with different concentrations and combinations of auxins, cytokinins and malt extract for primary callus induction. The best response for primary callus induction (90%) was obtained when MS medium was supplemented with 5 mg/l 2,4-D and 500 mg/l malt extract. Best results for embryogenic callus induction (80%) were obtained in C8 medium. The induction of somatic embryos was highest when MS medium was supplemented with 1 mg/l BAP and maturation of somatic embryos occurred when MS medium was supplemented with 5 mg/l 2,4-D and 1 mg/l BAP. Maximum plantlets were regenerated (92%) from the somatic embryos on half strength MS medium with no hormones. The plantlets were successfully acclimatized in different potting mixtures and highest survival rate (100%) was achieved in potting mixture containing sand and peat moss (2:1).

7年生马尾松组培苗试验林经营效益

通过收集7年生马尾松组培苗试验林胸径、树高、材积和产脂量数据,利用项目投资的财务效益评估手段对其经济效益进行分析研究,为丰富马尾松人工林高效培育技术体系提供试验依据。结果表明:在当前立地条件和经营措施下,组培苗林分林木胸径、树高、平均材积、蓄积量、出材量为15.5 cm、10.03 m、0.1008 m^(3)、151.20 m^(3)·hm^(-2)、91.5879 m^(3)·hm^(-2),分别比实生苗提高了27.0%、21.9%、89.5%、89.5%、94.9%,中径材林木数量占总株数比例提升了30个百分点,组培苗林分生长优势突出,材种规格获得大幅度提升。采脂后组培苗林分产值、净收益、净现值、内部收益率(IRR)为142869.1元·hm^(-2)、54283.4元·hm^(-2)、19802.4元·hm^(-2)、17.2%,分别比未采脂提高了130.9%、855.1%、331.5%、14.7个百分点,比实生苗林分提高了368.6%、475.8%、203.9%、26.6个百分点。7年生马尾松组培苗林分经济效益显著,在立地条件相似地区应用马尾松组培苗造林,可大幅度提高人工林经营效益。

马尾松组培苗幼林采脂试验分析

【目的】研究马尾松组培苗幼林产脂量与生长性状间的变化规律和松脂主要组分及含量特征,为马尾松早期利用和高产优质脂用原料林培育提供试验依据。【方法】以7年生马尾松组培苗人工林为研究对象,设置标准地开展采脂试验,测定胸径和树高,计算单株材积和产脂量,进行不同径级(树高级)间的产脂量比较分析,并拟合产脂量与径级和树高方程;通过气相色谱/质谱联用技术(GC/MS)检测松脂样品主要化学组分和含量。【结果】7年生马尾松组培苗幼林平均产脂量为15.9 g·d^(-1)·株^(-1)。不同径级产脂量间差异显著(P<0.05),随林木径级增大产脂量随之增加,呈一元线性关系(R^(2)=0.99);不同树高级产脂量随树高增大而增加,呈幂函数关系(R^(2)=0.98)。胸径、树高和材积与产脂量间相关性极显著(P<0.01),可以实现同步改良。7年生马尾松组培苗幼林单位面积可采脂林木数量和产脂量分别是同等条件下实生苗林分的1.9倍和1.8倍。在40%的强度下,采脂对林木生长的影响尚未表现出明显差异。兼顾产脂量和生长性状,马尾松组培苗幼林起始采脂径级以18 cm径级为宜。从组培苗林分松脂样品中共检出α-蒎烯、β-蒎烯、长叶烯、石竹烯、海松酸、山达海松酸、枞酸等单萜、倍半萜和二萜类化学物质22种,其主要组分及含量与常规经营中龄以上马尾松林分松脂基本一致。【结论】该立地条件下营造的7年生马尾松组培苗幼林产脂量、松脂品质已基本满足行业需求,在规范采脂的前提下可对其进行生产性采脂利用。采用马尾松组培苗造林在提早采脂和增加单位面积松脂产量方面具有较大的优势和潜力,为实现马尾松的早期化利用和松脂原料林的高质量培育提供了新的选择。

马尾松组培苗幼林胸径和树高分布特征

收集营造于广西派阳山林场的7年生马尾松组培苗试验林的胸径和树高生长数据,对林分胸径分布、树高分布、树高—胸径关系进行测定分析,利用数学模型分别进行拟合和回归模拟,求解模型参数并通过χ^(2)法进行检验,使用决定系数(R^(2))、均方根误差(RMSE)和F值进行综合评价,以筛选最优模型。结果表明:马尾松组培苗试验林的胸径偏度系数(sk)>0,胸径峰度系数(k)<0;树高偏度系数(sk)<0,树高峰度系数(k)<0;表明林分内林木竞争较为激烈,林木胸径和树高离散程度较大,可适度间伐对林分结构进行调整。试验林胸径分布和树高分布最优模型均为peal-reed模型,决定系数(R^(2))均在0.98以上,均方根误差(RMSE)为1.64~5.69,F值为62.57~684.68。在4个候选模型中,非线性模型h=1.3+d^(2)/(c0+c_(1)d+c^(2)d^(2))对林分胸径—树高关系模拟效果最好,其决定系数(R^(2))、均方根误差(RMSE)、F值分别为0.94、0.2711、65.85。筛选的3个模型对试验林胸径和树高生长变化具有较好的预测能力,可为相同立地条件和经营措施下马尾松无性系人工林经营管理提供参考。

不同质量浓度Cu^(2+)和Ag^(+)对大麦成熟胚组培特性的影响

【目的】探究不同质量浓度Cu^(2+)和Ag^(+)对大麦成熟胚组培特性的影响。【方法】以Bowman、北青7号和光头大麦的成熟胚为材料,研究不同质量浓度Cu^(2+)和Ag^(+)对大麦成熟胚愈伤诱导率、长根率、长芽率、褐化率、绿化率、再分化率和愈伤湿质量的影响。【结果】Bowman、北青7号和光头大麦成熟胚的7种组培特性间存在不同程度差异。与对照相比,0.75 mg/L Cu^(2+)可减少10.43%的成熟胚长根、7.48%的成熟胚长芽、8.24%的愈伤组织褐化、20.74 mg的愈伤湿质量;4.00 mg/L Ag^(+)可提高6.47%的愈伤组织诱导率,可减少9.08%的成熟胚长根、7.96%的成熟胚长芽和9.67%的愈伤褐化;0.75 mg/L Cu^(2+)和4.00 mg/L Ag^(+)对愈伤细胞转绿、愈伤增殖和再分化均无抑制。【结论】适宜质量浓度的Cu^(2+)和Ag^(+)可有效抑制大麦成熟胚长根、长芽和愈伤组织褐化,Ag+还可有效提高愈伤诱导率;在基于MS培养基的大麦成熟胚愈伤组织诱导和再分化培养中,Cu^(2+)和Ag^(+)的适宜质量浓度分别为0.75和4.00 mg/L。研究结果可为构建大麦成熟胚的愈伤诱导及其再分化技术体系提供理论支持。

红松愈伤组织增殖阶段生长与蔗糖代谢的关系

针对增殖培养中不同类型的红松(Pinus koraiensis)愈伤组织,采用形态学观察、生长曲线绘制、糖代谢的测定,对具有成胚能力差异的两类愈伤组织(胚性愈伤组织、非胚性愈伤组织)进行比较。结果表明:形态学上,胚性愈伤组织半透明,浅白或白色,结构疏松,表面有大量半透明突起,湿润有黏性,可稳定继代增殖培养;非胚性愈伤组织表面致密,白色逐渐转变成黄褐色。在体视显微镜下观察,胚性愈伤组织由许多胚细胞和单个胚柄细胞组成;而非胚性愈伤组织由核质浓密的圆形细胞单一或聚集组成。增殖培养过程中,红松胚性愈伤组织与非胚性愈伤组织的细胞生长曲线基本呈“S”形,胚性愈伤组织、非胚性愈伤组织生长率在15、20 d时开始下降;胚性愈伤组织与非胚性愈伤组织中可溶性糖(果糖、葡萄糖、蔗糖)质量分数差异极显著,蔗糖质量分数基本维持不变,葡萄糖和果糖质量分数变化明显,呈现先升高再降低的走向;参与蔗糖合成的蔗糖磷酸合酶活性变化与两种转化酶活性变化规律相似,而蔗糖合成酶活性基本维持不变;增殖阶段,胚性愈伤组织的可溶性糖质量分数、蔗糖代谢酶活性始终高于非胚性愈伤组织。

Establishment of Plant Regeneration System from Immature Embryos of Maize(Zea mays L.) Inbred Lines

[Objective] The aim was to explore the conditions of high frequency induction of embryonic callus and plant regeneration of maize. [Method] Immature embryos of maize inbred lines were used as explants to study the effect of different 2,4-D concentrations on the induction of callus,the effect of different 6-BA concentrations on the differentiation of test-tube plantlet,as well as the effect of different IBA concentrations on the rooting of test-tube plantlet. [Result] 2,4-D showed obvious effect on the induction of inducement rate of maize,and the optimum induction medium was：N6 ＋ 2 mg/L of 2,4-D ＋ 500 mg/L of CH ＋ 500 mg/L of Pro ＋10 mg/L of AgNO3; the optimum differentiation medium was：N6 ＋ 0.5 mg/L of BA ＋ 500 mg/L of Pro; the optimum for the rooting of test-tube plantlet was 1/2 MS ＋ 0.5 mg/L of IBA. [Conclusion] The study had provided basis for the genetic transformation of maize.

Optimization of plant regeneration system in vitro culture in wheat

Studies were carried out to establish an efficient regeneration system of three bread wheat cultivars. Results showed induction medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) had a higher plantlet regeneration frequency than Piclorm, with an average frequency of 54% in all treatments. Optimal condition for different genotypic rice was as following: induction medium (MSS 3AA/2) with 0.5 mg L-1 2,4-D, regeneration medium (R) with 0.01 mg L-1 2,4-D and 3 mg L-1 KT. The average regeneration frequency reached 83.3% under the condition. Correlation analysis showed that root differentiation, in different level, correlated with green spot regeneration, and with the number of regenerated plants per callus. No correlation was found between green spots regenerated and the numbers of plants regenerated per callus.

湖北花楸种胚离体培养与快繁技术研究

为建立湖北花楸组培快速繁育技术体系,以湖北花楸种胚为外植体,研究不同植物生长调节剂对种胚萌发、茎段增殖和芽苗生根的影响。结果表明,湖北花楸种胚在MS+0.5 mg/L 6-BA+0.05 mg/L IBA+30 g/L蔗糖+7 g/L卡拉胶中萌发并正常生长,平均萌发率可达95%,远高于未添加激素培养基组合的平均诱导率(5%),诱导培养基中的胚正常生长,30 d时可萌发出多片真叶和腋芽;带芽茎段转入MS+0.25 mg/L 6-BA+0.04 mg/L IBA+30 g/L蔗糖+7 g/L卡拉胶中增殖培养30 d,平均每茎段诱导出2.08个芽苗,增殖率490.5%;若带芽茎段转入MS+0.5 mg/L 6-BA+0.04 mg/L IBA+30 g/L蔗糖+7 g/L卡拉胶中增殖培养30 d,平均每茎段诱导出4.1个芽,增殖率291%;芽苗在MS+0.5 mg/L NAA+30 g/L蔗糖+7 g/L卡拉胶中培养30 d,生根率可达84%,芽苗基部通过愈伤再生方式诱导产生不定根,平均每芽苗诱导出4.41条根。

月季育种中胚培养技术的应用

通过采用组织培养技术,可实现月季胚的快速繁殖与优质品种的选育。文章详细介绍了月季胚培养的关键技术步骤,包括外植体的选择与处理、接种及培养方法、增殖阶段管理、壮苗培养与生根技术以及最终的移栽过程。胚培养技术可显著提高月季的繁殖效率,为月季新品种的快速培育提供有效途径。

玉米幼胚和成熟胚愈伤组织分化反应性比较

以鲜食玉米为主的13种基因型的成熟胚和幼胚为材料,对它们的愈伤组织诱导,继代培养和分化进行了系统研究。结果表明:玉米的成熟胚和幼胚愈伤诱导率相关,但两种胚诱导出的愈伤组织状态明显不同,幼胚诱导出的愈伤组织状态普遍比成熟胚的好;两种胚的愈伤分化率差异明显,成熟胚愈伤组织最高分化率为2.5%,幼胚愈伤组织最高分化率可达83.3%。

棉花组织培养中畸形胚的发生和转化

在棉花组织培养中,时常有畸形胚的发生。根据畸形胚的表型,可分为九类:联体胚、子叶畸形胚、胚轴畸形胚、单极胚、多极胚、无极胚、重新愈伤化胚、玻璃化胚和白化胚。畸形胚的发生频率很高,可达94.0％。培养时间和培养基组成等均影响畸形胚的发生。通过调整培养基中激素含量可有效地使畸形胚萌发转化为正常苗。

胚龄和2,4-D浓度对玉米自交系幼胚愈伤组织诱导率的影响

玉米幼胚愈伤组织的诱导由于受多种因素的影响使其相对于其它禾本科植物而言较为困难 ,其中胚龄和 2 ,4_D浓度是影响玉米幼胚愈伤组织诱导的两个较为主要的因素。本文分析了胚龄和 2 ,4_D浓度对玉米幼胚愈伤组织诱导率的影响 ,提出了玉米幼胚组织培养的最适的胚龄因自交系而异 ,对于吉 85 3,吉 842两种自交系胚龄以 11～ 13d为宜 ,而 4112自交系则以 12～ 14d为宜 ;3种自交系诱导的适宜 2 ,4_D浓度都为 1mg/L ,自交系间差异不大。为进一步研究能长期继代的玉米幼胚胚性愈伤组织的诱导 。

小麦幼胚的脱分化状态及再生性能研究

通过对30个品种(系)的研究,确立了小麦幼胚最佳取材时期的种子形态指标:在幼种子发育到嫩绿色,脊部由光亮转变为无光泽并具一层绒毛时,幼胚刚好发育到透明的后期和半透明前期。该时期仅有几个小时。探讨了不同基因型的幼胚的致密愈伤组织诱导状况及其继代保持和分化能力,并获得了良好的再生体系。使得致密愈伤组织诱导率达到91%,在14d时分化频率达到96 7%,70d时分化频率仍可达62%。并就温室材料和大田材料以及不同年份大田材料的致密愈伤组织的继代保持和分化能力进行了比较。

枣胚培技术体系的建立

以‘六月鲜’枣为试材,对花后12～82d的幼胚进行培养。结果表明,适宜于胚挽救的最小胚龄为花后50d左右;在胚形成前进行离体培养,蔗糖浓度以7%为宜,而胚形成后离体培养以5%蔗糖为宜;暗培养可促进离体胚的生长发育,且有利于生根;植物生长调节剂及有机添加物极显著地影响着胚培效果。花后70～72d胚龄的胚适宜培养基为1/2MS+IBA0.4(mg/L,单位下同)+BA0.8+LH500+GA35+5%蔗糖。胚培苗经扩繁后转入1/2MS+IBA1.0+IAA0.06+2%蔗糖培养基上,获得了健壮生根苗,生根苗移栽成活率达85%以上。