Efficient In Vitro Shoot-regeneration Systems in Cassava (Manihot esculenta Crantz)

Two different protocols for in vitro regeneration of cassava using zygotic embryos and nodal axillary meristems have been developed. In both cases, buds were regenerated directly from excised explants without an intervening callus phase after a two-step culture procedure. In cotyledonary explants derived from zygotic embryos, prolific shoot formation occurred within 2—3 weeks on MS medium supplemented with 0.5—5 mg/1 BAP alone or in combination with 0.1 mg/1 NAA. Nodal explants with axillary meristems derived from aseptically grown seedlings or stem cuttings were used to initiate a round compact bulb-like structure on MS medium containing 10 mg/1 BAP. These latter structures, when cultured on MS medium supplemented with 0.1 mg/1 NAA, 1 mg/1 BAP and 0.1 mg/1 GA₃, produced multiple shoots. Somatic embryos isolated at the globular/torpedo stage from zygotic embryo explants were also capable of multiple shoot production on medium with 1 mg/1 BAP. Rooting of regenerated shoots exceeded 95 % in phytohormone-free MS medium. No change in their ploidy levels was observed. Therefore, the protocols developed should be of use in the particle gun and Agrobacterium-mediated genetic transformation of cassava.     

山羊卵泡卵母细胞体外成熟的研究

为了建立针对山羊卵母细胞的理想体外成熟培养体系,对山羊卵泡卵母细胞的采集方法,FSH和LH对山羊卵泡卵母细胞体外成熟的作用,及培养时间对卵母细胞体外成熟的影响进行了研究。结果表明:采用切割法平均每枚卵巢采集到的A,B级卵母细胞的数量明显多于抽吸法;采用TCM199添加体积分数为10?S,1~2 mg/L 17β_E2,10 mmol/L Hepes,0.12 mg/mL丙酮酸钠,0.1 mg/mL链霉素和0.125 mg/mL青霉素,10 mg/L FSH及20~50 mg/L LH的培养体系,有利于山羊卵母细胞的体外成熟,并放出第一极体。山羊卵母细胞体外成熟培养20,22,24 h,成熟率显著高于培养18,26,30 h(P<0.05)。     

甘蓝型黄籽油菜子叶离体培养初报

以甘蓝型油菜黄籽双单倍体品系04K91为供试材料,以无菌苗子叶为离体培养外植体,考察了苗龄（7d、10d、13d）与外植体部位及大小（带柄子叶、1/4子叶片、1/8子叶片和子叶柄切段）对芽再生频率的影响。方差分析结果表明,苗龄对芽再生频率有显著影响,外植体对芽再生频率有极显著影响,而苗龄×外植体对芽再生频率影响不显著。多重比较结果表明,以7d苗龄无菌苗的带柄子叶或1/4子叶片为外植体较好,出芽率较高,达90%以上。     

大豆尿黑酸叶绿基转移酶基因的克隆与进化分析

采用基于EST的电子克隆方法,以拟南芥(Arabidopsis thaliana)的尿黑酸叶绿基转移酶基因(Homogentisate phytyltransferase,HPT)cDNA序列为信息探针,对大豆的EST数据库进行同源检索筛选,获得了1 777 bp长大豆HPT基因的cDNA序列(GenBank登录号为:AY956421),经RT-PCR扩增、分子克隆和序列分析验证,表明与电子克隆序列一致;该基因具有完整的开放阅读框架(ORF,173～1 408 bp),推测编码411个氨基酸的蛋白。该蛋白与拟南芥(Ara-bidopsis thaliana)、苜蓿(Medicago sativa)、水稻(Oryza sativa)、玉米(Zea mays)、光合集胞蓝细菌(Synechocystis)的序列进行了比较,发现该基因具有保守性。     

Evaluation of Near Infra-red Reflectance Spectroscopy for Predicting Grain and Stover Quality Traits in Maize

The prediction of grain and stover quality parameters in maize {Zea mays L.) by near infra-red reflectance spectroscopy (NIRS) was studied. A total of 110 grain and 135 stover samples originating from different genotypes and environments were assayed. Calibration equations for content of crude protein (CP), crude fat (CF), starch (ST), and water soluble carbohydrates (WSC) in grain were obtained by multiple linear regression of known manual values on NIRS data from the odd numbered samples. Calibrations for CP, acid detergent fibre (ADF), in vitro digestible organic matter according to the Tilley & Terry (IVDOM-T & T) and the gas production (IVDOM-Gp) method, respectively, and metabolizable energy (ME) in stover were developed analogously. Equations were validated with the evennumbered .samples and for ME additionally with the 1584 stover samples from an experiment with 66 F₁ hybrids tested in six environments. The coefficients of multiple determination (R²) of the prediction equations ranged from 0.80 for IVDOM-Gp and ME in stover to 0.94 for CP in grain. Standard errors of calibration (SEC) and prediction (SEP) were in most cases not higher than commonly reported for conventional manual assays. With regard to the correct ranking of hybrids, prediction equations for ME applied well to stover samples from other environments with one exception. We concluded that NIRS can evaluate the quality traits investigated to a similar degree to that of conventional methods of analysis. Since NIRS is simple and safe to operate and allows rapid screening of several quality traits simultaneously, it should be particularly attractive for breeding purposes.     

In vitro pollen germination and tube growth of tomato (Lycopersicon esculentum Mill.) and its relation with plant growth

Summary In vitro pollen germination at 10°C, 14°C and 22°C of four groups of two pure line tomato varieties was compared with their plant growth at 19°C D/10°C N under controlled environmental conditions. Generally, pollen germination was slow at 10°C but after 6 h the percentage of germination was similar to that at 22°C. Maximum germination was obtained at 14°C already after 4 h. The longest pollen tubes occurred at 22°C. The two varieties within each group differed significantly from each other for percentage of pollen germination. For one group, this difference was greater at 10°C than at 22°C. The varieties in two groups also differed significantly for pollen tube growth. These differences in pollen tube growth rate were greater at 22°C than at 10°C. There was no clear relationship between pollen germination, pollen tube growth and plant growth in any of the four pairs of varieties. The results are discussed with regard to the possibility of pollen selection at low temperature in order to improve the efficiency of breeding for growth at low temperature.     

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.     

葡萄试管苗土支撑培养带坨移栽研究

以土作为培养基支撑物，离体培养葡萄试管苗双节茎段，４０天后即达到适于炼苗标准，成苗率９５．８％。带坨移入营养钵，在不喷雾，不覆膜，空气湿度低至４５％条件下，成活率达９６．７％。成活的试管苗炼苗５４天后移入大田，移栽成活率为１００％，其中特级苗占８７．５％，一级苗占１２．５％。试验证明，土支撑培养带坨移栽技术是简化试管苗移栽程序，降低成本的有效措施。     

穗离体培养下温度对小麦蛋白质及其组分和产量性状的影响

本实验采用穗离体培养的方法研究了不同培养温度对小麦蛋白质及其组分和小麦产量性状的影响.试验表明:在扬花前3天至乳熟期间,温度对小麦籽粒总蛋白质的含量影响不大,但对蛋白质组分影响较大,低温有利于清蛋白的形成,高温有利于醇溶蛋白和谷蛋白的形成;同时试验还表明低温条件下小麦茎杆总蛋白含量高于高温培养下的茎杆含量;不同培养温度对小麦结实率和粒重也有影响,小麦1.2小花和整穗可见花结实率随温度增高而上升,到20℃时达到最大,分别为96.36%和72.11%,百粒重随温度的升高而增大,呈线性相关,相关系数为0.974013.另外本试验还对受温度影响的13个指标进行了因子分析,分析结果表明13个指标可以压缩为3个因子,因子1可以命名为产量因子,决定总变异的48.045 4%,因子2和因子3可合称为蛋白质组分和粒重因子决定总变异的51.9546%.     

Mature plant and tissue resistance in the groundnut-peanut bud necrosis virus system

Summary Leaves and plants of different ages of a susceptible and two resistant groundnut genotypes were mechanically inoculated with peanut bud necrosis virus, and the percentage of plants with systemic symptoms (incidence) and the incubation period were determined. The incidence decreased sharply in all three genotypes with the age of the inoculated leaves and plants. The incubation period increased with the age of leaves and plants. Apparently, only young tissue of young plants is susceptible, while mature tissue and plants are highly resistant. This mature tissue and plant resistance occurs irrespective of the susceptibility level of the genotype to peanut bud necrosis virus, however, it develops earlier in the resistant than in the susceptible genotypes.Abbreviations IP₅₀ incubation period - PBNV peanut bud necrosis virus - TSWV tomato spotted wilt virus