甘蓝型黄籽油菜原生质体的游离和纯化

以西南农业大学选育的渝黄1号和SH21 2个甘蓝型黄籽油菜品种为材料,切取其无菌苗的子叶和下胚轴分离纯化原生质体。种子萌发时低温处理可明显提高原生质体活力;光照处理对子叶原生质体活力有较大影响;酶液配比对原生质体产率影响较大;用“过滤－离心－漂浮法”进行纯化,用21%蔗糖,800转/min纯化效果较好。     

一例猪繁殖与呼吸综合征的RT-PCR诊断与防制

2008年3月,新乡市某猪场发生了以一种经产母猪和初产母猪流产、死产、产木乃伊胎、死胎及弱仔为特征的疾病。采集流产胎儿和死胎的肺、肝、脾脏等病料,制悬液,取上清,进行细胞培养,观察细胞病变。另根据猪繁殖与呼吸综合征病毒美洲株VR-2332 ORF7基因序列设计一对特异引物,提取流产胎儿的肺、肝等组织中PRRSV的RNA,经RT-PCR扩增,检测猪繁殖与呼吸综合征病毒为阳性,结合病史和临床症状,诊断为猪繁殖与呼吸综合征。通过对该猪场采取紧急接种疫苗、彻底消毒等综合防制措施,控制了该病的流行与蔓延。     

洋葱愈伤原生质体分离和纯化研究

建立高效原生质体分离和植株再生体系是进行体细胞杂交的基础性工作。以洋葱多次继代的愈伤组织为研究试材,对原生质体分离纯化技术进行研究。结果表明：2.0%纤维素酶Onozuka R-10+0.1%果胶酶Y-23+0.5%离析酶 R-10+0.60 mol/L甘露醇+CPW+0.1% MES的酶液,酶解6 h,洋葱原生质体分离效果最好。纯化时,适当降低离心力的短时间离心有利于原生质体纯化过程中产量和活力的保持,以600 r/min离心5 min效果为好。     

玉米、绿豆原生质体的游离和融合

现代科学技术以细胞融合为手段,创造作物新品种已成为可能.我们采用玉米叶肉细胞和绿豆根尖细胞分离出原生质体取得成功.将二者进行融合也取得了一定进展.     

锰矿废弃地几种不同恢复模式土壤种子库与地上植被的关系

土壤种子库技术是矿区废弃地植被恢复的重要方法,弄清不同恢复模式下种子库与地上植被之间的关系是有效利用该技术的前提。以云南建水锰尾矿恢复的醉鱼草(Buddleja officinalis)灌丛、香根草(Vetiveria zizanioides)草丛、肿柄菊(Tithonia diversifolia)群落为对象,以人工林蓝桉(Eucalyptus globulus)林为对照,通过野外调查与室内萌发实验相结合的方法研究了各群落地上植被与土壤种子库的关系。结果显示:1)尾矿起源各群落地上植被(0.50、0.06、2.18、vs. 2.12)与土壤种子库物种多样性(1.32、1.42、1.78 vs. 2.65)均低于对照群落;2)地上植被物种多样性与土壤种子库物种多样性成正相关(R^2=0.187,p=0.005);3)对照群落土壤种子库与地上植被物种相似性较高,尾矿恢复各群落较低,且不同恢复模式间相似性差异较大;4)香薷(Elsholtzia ciliata)与醉鱼草为4个群落土壤种子库共有的优势种,各群落地上植被与土壤种子库优势种不完全同步。研究结果表明恢复措施对土壤种子库物种多样性有重要影响。     

三里洞煤矸石废弃地景观规划

煤矸石的大量堆放破坏环境与景观格局,带来了一系列的生态环境问题。本文探索了矸石废弃地景观规划设计的目标与原则,运用生态恢复、景观设计的理论方法指导矸石废弃地进行生态恢复与景观重建。以矸石堆尺度、废弃地尺度两种立地类型划分结果为依据,在此基础上进行了景观规划设计。从而引发对废弃地区域规划设计的思考,这对改善矿区生态环境面貌,恢复土地的生产能力,促进农业生产,提高城市地位和形象有着重要的意义,也可为其它地区矸石废弃地的景观规划提供借鉴。     

新牧4号紫花苜蓿原生质体游离和培养条件的研究

主要探讨影响苜蓿原生质体游离和培养的条件,为原生质体的培养体系的建立提供了依据。以‘新牧4号紫花苜蓿’的子叶、下胚轴和愈伤组织为材料,研究了酶液的pH值,酶解时间,酶液组合和不同培养方法对原生质体游离和培养效果的影响。结果表明：当酶液pH值为5.8时,有活力的原生质体的产量最高,同时细胞碎片少;子叶、下胚轴和愈伤组织适宜的酶解时间均为10 h;酶液组合为2%纤维素酶＋0.5%果胶酶＋0.3%或0.5%离析酶,游离出的有活力的原生质体产量较高;采用液体浅层培养和固液双层培养,均观察到原生质体发生分裂,但固液双层培养法更有利于‘新牧4号紫花苜蓿’原生质体的分裂和培养。     

宁南黄土丘陵区不同生态恢复与重建中的土壤水分变化研究

针对宁南黄土丘陵区存在的生态脆弱、环境恶化的突出问题,对黄土丘陵区不同生态恢复过程中的土壤水分变化进行了研究。研究结果表明：在退化山地的植被恢复过程中, 0~60cm土层,土壤水分含量随恢复年限的增长而增加,在60cm土层以下,土壤含水量随着退耕年限的不断增长不断减小;对于不同恢复方式的土壤水分含量,88542水平沟>人工草地>鱼鳞坑>天然草地;对于不同植被类型的土壤水分含量,草地>灌木地>林地;对于不同地形的土壤水分含量,阴坡>半阴坡>阳坡,坡下部>坡中部>坡上部,并且随着坡度的不断增加,土壤水分含量不断减小。     

Genetic evaluation of cultivated garlic germplasm (Allium sativum L. and A. ampeloprasum L.)

Garlic cultivated in rural farms of South Italy is often a heterogeneous clone population, which can comprise different cytotypes. A collection of cultivated garlic from the University of Basilicata gene-bank, was evaluated for ploidy level, and 16 different morphological traits. Out of50 accessions, 7 were hexaploids (Allium ampeloprasum L.) and43 diploids (Allium sativum L.). Significant differences in yield were observed within and between ploidy levels. The heritable traits most correlated with yield were plant height (h² = 0.62) within diploid types and neck diameter (h² = 0.75) within hexaploids. Discriminant analysis did show that four characters (leaf basal width, total n^° of leaves, clove diameter and neck height) were able to correctly discriminate all germ plasm accessions between the two species. This revised version was published online in August 2006 with corrections to the Cover Date.     

乐都紫皮大蒜叶片生长发育与果聚糖代谢的关系

为了探讨高原大蒜生长特性及其果聚糖代谢规律,分析叶片生长发育与果聚糖代谢的关系,本研究以"乐都紫皮大蒜"为材料,对不同发育期的大蒜叶片生长情况、生理指标及其果聚糖代谢关键基因的表达特征进行了解析。结果表明:随着生育期的延续,大蒜植株的最大叶长、最大叶宽和单株叶片数均稳步增加。叶片生理指标呈现先升高后降低的趋势,总叶绿素含量在花芽分化之前快速上升,在抽薹期达到最大值2.32 mg/g FW;组织含水量在鳞茎膨大初期达到最大值89.06%,随后急剧降低。在整个发育过程中,大蒜叶片蔗糖:蔗糖1-果糖基转移酶基因(1-SST)和果聚糖外切水解酶基因(1-FEH)表达量存在显著差异,从幼苗期至鳞茎膨大初期,1-SST基因表达量显著高于1-FEH。1-SST的表达在全生育期表现为"升高-降低"的趋势,在抽薹期,1-SST的表达量达到峰值,比幼苗期提高了17.51倍;1-FEH在全生育期的表达量均较低,呈现起伏波动的趋势,在收获期表达量最高,是苗期的2.41倍。综合分析说明,大蒜叶片叶绿素和果聚糖的合成高峰主要在抽薹期,抽薹期是大蒜叶片生长发育和果聚糖代谢的关键时期,大蒜可通过叶绿素的合成以及1-SST和1-FEH的差异表达进一步影响鳞茎果聚糖的累积。     

Chromosome and cytoplasm analyses of somatic hybrids between onion (Allium cepa L.) and garlic (A. sativum L.)

Chromosomes and cytoplasms were analyzed in two lines of a somatic hybrid between onion (Allium cepa L.) and garlic (A. sativum L.). One line of the somatic hybrid had 40 chromosomes and the other 41chromosomes. Genomic in situhybridization successfully revealed the chromosome constitution of the two lines. One line had 20 chromosomes from onion and17 chromosomes from garlic, and the other had 21 chromosomes from onion and 17chromosomes from garlic. Interestingly, both lines had three chimeric chromosomes. PCR-RFLP analyses of chloroplast and mitochondrial DNAs of both lines showed that these were identical to the onion parent. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity of garlic (Allium sativum L.) germplasm from China by fluorescent‐based AFLP,SSR and InDel markers

Garlic (Allium sativum L.), an asexually propagated crop, is an important vegetable and medicinal plant. China is the biggest garlic producer in the world; however, the genetic background of garlic from China is not well understood. In this study, population structure and clustering analysis of garlic germplasm was performed using amplified fragment length polymorphism (AFLP), simple sequence repeat (SSR) and insertion–deletion (InDel) markers. Among 212 accessions of garlic, genetic diversity analysis identified 546 alleles amplified by AFLP, SSR and InDel primers, and 492 of these were polymorphic. All accessions were divided into five groups by structure analysis and neighbor‐joining clustering. Most traits, including allicin content, were only slightly affected by population structure, which indicated that this germplasm can be used as populations for association mapping. The results provide a molecular basis for understanding the genetic diversity of the garlic germplasm preserved in China.     

植物根际促生菌对大蒜的促生、抗病作用研究

为了研究不同剂量的植物根际促生菌(PGPR)对大蒜生长时期的促生和抗病作用,以盆栽种植的大蒜种子为试材,用计数法计算其发芽率和常规测量法测定其株高;再用土壤温湿度检测仪测定种植所用土壤养分含量。与CK组相比,实验组B（施用4 g菌肥）铵态氮提高117.60%,有效钾提高593.15%,实验组C（施用6 g菌肥）速效磷提高238.77%;而实验组C的大蒜的发芽率提高15.17%,幼苗株高提高20%,叶绿素含量提高11.04% (P<0.05),这说明了PGPR对大蒜生长有促进作用。大蒜根腐病菌与植物根际促生菌的拮抗试验结果表明在30℃暗培养下,菌肥浓度为10^-2时有害微生物指数下降25%,表明其抑菌效果显著。本研究结果表明PGPR对大蒜促生作用显著,同时具有一定抗病效果,在农业发展中具有潜在的经济价值。     

The induction of tetraploidy in chicory (Cichorium intybus L. var. Magdebourg) by protoplast fusion

Summary Fusions of chicory leaf protoplasts were realized in order to obtain a higher number of tetraploid plants compared to the traditional doubling techniques with colchicine. Thanks to the fusion technique, using a mixture of PEG, DMSO and a solution of CaCl₂ (pH 10.5), 25% tetraploid (2n=36) plants were obtained. Among the 167 regenerated plants, only one aneuploid of a near tetraploid level (2n=33) was identified.     

Genetic diversity among selected Argentinean garlic clones (Allium sativum L.) using AFLP (Amplified Fragment Length Polymorphism)

Genetic diversity of eight selected Argentinean garlic clones (Allium sativum L.) were investigated at the DNA level with the amplified fragment length polymorphism DNA (AFLP) procedure. A total of 405 unambiguous bands were identified by six primer combinations of EcoR I +3 and Mse I +3, of those, 398 showed a clear polymorphism, representing 98% of the total bands. A presence/absence matrix was constructed with the polymorphic bands, and a dendrogram was obtained from it with the UPGMA method. The accessions showed different levels of similarity ranging between 0.24 and 0.97, using the coefficient of Jaccard. The dendrogram showed six arbitrary groups. Accessions typically considered as different clones show similarities between 0.97 and 0.495. The garlic clones were clustered according to the physiological group and bulb color. We could detect an association between AFLP and the geographical origin of the clones. The potential use of AFLP could allow not only the differentiation among species, but also between botanical varieties and well-defined ecotype groups. This is the first report of the use of AFLP to characterize Argentinean garlic clones. This revised version was published online in August 2006 with corrections to the Cover Date.     

茶树叶片和胚根原生质体的分离及PEG诱导融合

植物原生质体是细胞培养和体细胞融合等细胞水平研究及植物遗传育种的重要材料。本研究用福鼎大白茶茶树的幼嫩叶片及胚根, 分析了原生质体分离过程中的材料、酶解液组成及酶解时间、纯化方法等影响因子, 建立了最佳原生质体分离体系, 为茶树体细胞杂交等细胞水平的研究提供了高效获取大量高活力原生质体的方法。结果表明, 23°C恒温黑暗或遮光培养的茶树实生苗的5周叶龄以内的幼嫩叶片是茶树原生质体分离的最佳材料, 其次是茶树种子萌发后的幼嫩胚根; 而以茶园健康生长的5周叶龄以内的幼嫩叶片为材料时, 只能获得混有大量细胞碎片的少量具有活力的原生质体。以茶树幼嫩叶片为分离材料的酶解液组成为1.5%纤维素酶+0.1%离析酶+0.5%果胶酶+0.4 mol L^-1甘露醇+20 mmol L^-1 MES; 以茶树幼嫩胚根为分离材料的酶解液组成为1.5%纤维素酶+0.3%离析酶+0.5%果胶酶+0.4 mol L^-1甘露醇+20 mmol L^-1 MES。分离茶树幼嫩叶和幼嫩胚根原生质体时, 宜采用低速(分别为55 r min^-1和50 r min^-1)恒温(23°C)摇床振荡酶解培养, 时间分别为7 h和8 h; 最适宜采用15×g的转速, 离心4 min可纯化获得高产量和活力的原生质体。用40% PEG-6000诱导20 min后可使茶树原生质体融合, 融合率达10%。     

Synthesis of low linolenic acid rapeseed (Brassica napus L.) through protoplast fusion

Brassica napus somatic hybrids with low linolenic acid (18:3) content in their seed oil have been produced using fusion partners screened for low 18:3. One somatic hybrid contained only 3.5% 18:3, a level significantly below the mid-parental mean. The low level of 18:3 proved stable in the R₁ generation. Oil content of the lowest 18:3 selection increased from the mid-parental mean (29.3%) in the R₀ generation to 36% in a R₁ field bulk. The R₁ field population also showed some resistance to shattering. This revised version was published online in July 2006 with corrections to the Cover Date.