犬造血干细胞移植的作用机制及应用研究进展

造血干细胞(hematopoietic stem cell, HSC)位于造血系统的顶端, 是造血系统中唯一具有多能性和自我更新能力的细胞, 可以分化为各种功能的血细胞, 维持血液系统的建立和动态平衡, 是目前研究最为透彻、临床应用最为成熟的成体干细胞。造血干细胞的这些重要特性, 使得基于造血干细胞在治病机制研究和临床应用中的研究取得了很大进展。以造血干细胞为基础的再生医学治疗是目前治疗恶性血液病和遗传病的首选方法, 如造血干细胞治疗犬白细胞黏附缺陷综合征、犬遗传性贫血、犬淋巴瘤、犬白细胞减少症、犬X连锁严重联合免疫缺陷病等, 但由于白细胞抗原匹配的供体稀缺、可获得的造血干细胞数量有限等原因, 无法满足临床需求, 因此, 如何通过体外培养获得满足临床需要的造血干细胞成为近年来学者们研究的热点问题。作者参照现有研究成果对造血干细胞的来源和体外分离培养、治病机制进行系统的描述, 并对造血干细胞移植治疗遗传性溶血性贫血、白细胞黏附缺陷综合征、淋巴瘤、白细胞减少症、X连锁严重联合免疫缺陷病的临床研究进行综述, 以期为今后将造血干细胞广泛应用于临床治疗提供参考依据。     

观赏羽衣甘蓝的离体繁殖

以花蕾为外植体，离体繁殖羽衣甘蓝８个杂各一代材料。培养基为（ｍｇ／Ｌ）：ＭＳ＋ＢＡ１－２＋ＮＡＡ０．１－０．２＋ＩＡＡ０．５＋（或不加）ＧＡ３０．３＋３％蔗糖。平均１８％－６７％的接咎花蕾产生不定芽；药有３０％的不定芽发育成完整植物。培养物的严重褐化和再生植物的普遍玻璃质化是影响植株成活的主要因素。培养基中加入活性炭和青霉素，并给以强光照射，可提高植株成活率。     

山羊卵泡卵母细胞的采集及体外成熟效果研究

为了获取高质量的卵泡卵母细胞，提高山羊体外胚胎生产的效率，采用切剖法与抽吸法采集卵泡卵母细胞，并对获得的可用卵母细胞进行了体外成熟培养。结果表明，切剖法能够获得较多的可用卵母细胞，可以明显增加用于体外成熟的卵母细胞数；卵母细胞的体外成熟率与卵泡直径密切相关，大卵泡与中卵泡的卵母细胞成熟率明显高于(P<0.05)小卵泡；山羊卵泡卵母细胞的体外成熟培养液中FCS的添加比例以10% ~ 15%为益。在一定范围内，提高FCS的浓度，并不能提高卵母细胞的成熟率。     

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。     

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.     

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     

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.     

Mutation breeding of Chrysanthemum morifolium Ram. using in vivo and in vitro adventitious bud techniques

Summary During experiments, which are being carried out to study the factors which control the process of adventitious bud formation in vivo on detached leaves of Chrysanthemum morifolium Ram, adventitious shoots were produced from leaves, irradiated with 500 rad of X-rays. The most important but disadvantageous result was that the majority of the adventitious shoots proved to be of a chimeral nature and obviously developed from more than one cell.An in vitro adventitious bud technique was developed using different types of explants. Pedicel segments regenerated the highest number of adventitious shoots and, moreover, they developed faster as compared to explants of young flower heads or leaves. The mutants produced by irradiating the various explants were almost exclusively of a solid (non-chimeral) nature. In addition, histological observations suggest that single epidermal cells are involved in the initiation of the adventitious shoot apices.The optimum dose for mutant production is approximately 800 rad X-rays. Rather often, more than one phenotypically identical mutant was found, which was always derived from the same explant. They could for instance originate from a multi-apical meristem formed by a single mutated cell.