尼罗罗非鱼精巢支持细胞分离培养体系建立及优化

本研究分离培养尼罗罗非鱼(Oreochromis niloticus)精巢支持细胞(Sertoli cells, SCs),建立并优化尼罗罗非鱼精巢支持细胞分离培养体系。无菌条件下,取发育至第Ⅲ期的尼罗罗非鱼精巢,PBS清洗后,剪碎精巢组织,0.25%胰蛋白酶消化,用含10%犊牛血清(Newborn bovine serum, NBS)的L-15培养液终止消化,过滤、离心,获得细胞。差速贴壁法获得SCs后,在26℃、无CO2饱和湿度恒温培养箱中培养。分别采用含10% NBS的L-15、M199、F12培养液,含5%、10%、15% NBS的L-15培养液,含1%罗非鱼血清的L-15+10% NBS培养液培养尼罗罗非鱼SCs。各组每2 d取细胞计数,绘制SCs生长曲线;甲基绿染色,倒置显微镜下观察SCs的形态。尼罗罗非鱼SCs培养1~2 d,细胞贴壁;培养3~5 d,细胞完全贴壁并迅速增殖。单个SCs呈不规则多边形,其核位于胞质中央,呈卵圆形,胞质中可见吞噬颗粒和空泡,空泡聚集于支持细胞胞质的两极或散布于核的四周。SCs在L-15培养液中贴壁生长,在F12、M199培养基中较难贴壁。与F12、M199培养液相比,L-15培养液更有助于尼罗罗非鱼SCs生长增殖(P<0.01)。与添加5%和15% NBS相比,在L-15培养基中添加10% NBS更有助于尼罗罗非鱼SCs生长增殖(P<0.05)。与未添加尼罗罗非鱼血清相比,在10% NBS+L-15培养中添加1%尼罗罗非鱼血清,能显著促进SCs生长增殖(P<0.05)。研究表明,采用胰酶消化差速贴壁法获得尼罗罗非鱼精巢支持细胞,10% NBS+1%罗非鱼血清+L-15培养液培养,能显著促进尼罗罗非鱼精巢支持细胞生长增殖。     

利用游离小孢子培养育成早熟春甘蓝新品种‘豫生1 号’

 采用游离小孢子培养技术快速育成早熟春甘蓝一代杂交新品种‘豫生1 号’ ( 96-1) 。该品种叶球近圆形, 结球紧实, 叶质脆嫩, 耐先期抽薹, 耐裂球, 抗病毒病、黑腐病、霜霉病。定植后60 d 成熟, 净菜产量44~ 55 t/ hm²。适于华北地区春季栽培。     

芥蓝游离小孢子培养初报

 以芥蓝的5个基因型为试材，对游离小孢子培养诱导胚状体发生及植株再生进行了初步探讨。结果表明：供体基因型对成功诱导胚状体发生至关重要；振荡培养明显有利于胚状体的发育；改良MS和B培养基对诱导植株再生没有差别。     

利用游离小孢子培养技术育成早熟大白菜新品种‘豫新60’

 ‘豫新60’系采用游离小孢子培养双单倍体育种技术快速育成的大白菜一代杂种。该品种早熟, 生育期60～65 d ; 高抗病毒病( TuMV) 、霜霉病和软腐病; 净菜产量59.9～69.8 t·hm^-2; 叶球矮桩叠抱, 球形指数1.19 , 单球质量1.5～2.0 kg ; 叶质柔嫩, 生食、熟食皆宜; 口感略甜, 风味佳。     

结球甘蓝小孢子培养条件优化及高代自交系胚状体诱导研究

以25份不同类型的结球甘蓝为试验材料,研究游离小孢子培养过程中影响胚状体产生的因素并进行了优化;同时对结球甘蓝高代自交系的诱导出胚情况进行了研究。结果表明:基因型是影响结球甘蓝胚状体产生的关键因素,参试材料中有11份(2份一代杂种和9份高代自交系)出胚,其中一代杂种中中甘628出胚率(19.8个·蕾~(-1))最高,高代自交系中01-88出胚率(47.5个·蕾~(-1))最高,极显著高于其他参试材料;花蕾长度3.0~3.5 mm、花药长度∶花瓣长度为3∶2至2∶1时,处于单核靠边期的小孢子比例最高,是适合小孢子培养的最佳取样时期;初花期和盛花期是理想的取蕾时期;添加适量活性炭可以极显著提高胚诱导率。     

红菜薹游离小孢子培养技术研究

对影响红菜薹游离小孢子培养的关键因素进行分析。研究结果表明,不同红菜薹材料之间的小孢子胚诱导率差异很大;4℃低温预处理2h对小孢子胚的形成具有促进作用;NLN培养基中添加生长素(NAA)和细胞分裂素(6-BA)对小孢子胚诱导率影响不大,添加浓度过大时诱导率反而降低;培养基中添加活性炭能提高小孢子胚诱导率;子叶型胚在15g/L琼脂的培养基上直接成苗率最高,达56.7%;再生植株在MS和1/2MS IBA(0.5mg/L)中的生根能力最好,1/2MS生根效果其次。     

Association of vegetation and soil mite assemblages with isolated Scots pine trees on a Scottish wet heath

Isolated trees may significantly enhance biodiversity at the landscape level. However, our understanding of their impacts is still poor, particularly in environments with high soil moisture where research on this topic has been comparatively limited. We examined understorey vegetation and soil oribatid mite assemblages under live and dead Scots pine trees and in open treeless areas, all within the same Scottish upland wet heath system, to determine whether isolated live trees affected the understorey and mite components of the ecosystem, and whether these effects occurred in parallel. We also explored whether these responses might result from tree-driven reductions in soil moisture content. Live trees reduced soil moisture (relative to wet heath and beneath dead trees) and appeared to change vegetation from wet heath to dry heath type communities. These effects were strongly related to tree trunk diameter (tree size). No major effects of dead trees on understorey vegetation or soil moisture were apparent. Higher mite species abundance and richness were found under live trees than in treeless open heath. Although mite abundances were lower under dead trees than live trees, richness remained similar, thus different factors seem to be regulating mite abundance and community composition. These findings indicate that landscape-level biodiversity responses to environmental change such as habitat fragmentation cannot be predicted from vegetation patterns alone, and that even in heavily fragmented landscapes comparatively small patches such as isolated individual trees can enhance biodiversity.     

大白菜游离小孢子培养和植株再生

以4个福山包头系列大白菜品种为试材,对游离小孢子培养和植株再生过程中的一些重要因素进行了研究。结果表明,不同基因型对小孢子胚诱导率和成苗率影响较大,平均每个花蕾产胚在1.28~38.25个之间;且胚胎发生能力高的品种,成苗率较高,胚胎发生能力低的品种,成苗率也较低;培养基中添加0.1~0.3 g/L活性炭能明显提高小孢子胚诱导率;及时将成熟的子叶形胚转移至再生培养基上,对提高成苗率至关重要;琼脂浓度为1.0%~2.0%时,成苗率大大提高。     

赣优3号苦瓜离体快繁体系的建立

研究了不同浓度生长素和细胞分裂素配比对"赣优3号"苦瓜丛芽诱导、增殖和生根的影响,获得了增殖倍数高、根系发达的完整植株。农艺性状评价表明,组培苗与实生苗在植株和果实形态、商品产量等方面无明显差异。本研究初步建立了"赣优3号"苦瓜的离体快速繁殖体系。     

小白菜小孢子胚状体诱导影响因素研究

以10个小白菜杂交种为试材,采用游离小孢子培养方法,研究影响小孢子胚状体形成因素的结果表明,不同品种胚状体的诱导率不同;NLN培养基中大量元素减半有利于小孢子胚诱导,其诱导率达到100%;培养基中添加0.05 mg/L的6-BA和0.2 mg/L的NAA可以提高胚状体诱导率;活性炭和琼脂糖对诱导胚状体发生和发育有促进作用,可使子叶胚的比例达90%以上;花蕾经4℃低温预处理24 h后的成胚率显著提高。