Chromosomal monogenic dominant male sterility in chinese cabbage (Brassica rapa supbs. pekinensis (Lour.) hanelt)

Summary Strong indication was found for the existence of a chromosal monogenic dominant male sterility-gene in chenese cabbage. This source of male sterility can be of practical use for the production of hybrid varieties. A pronounced drawback is the required removal, in the breeding and seed production stage, of the approximately 50% male fertile plants from each offspring of male sterile plants. Fortunately this removal is facilitated by the penomenon of apical dominace in the flowering process in the whole Brassica rapa taxon. Moreover an advantage of dominant monogenic male sterility is its easy and rapid introduction into all available genotypes of chinese cabbage.     

Effect of Calcium and Potassium Nutrition on Yield,Ion Content,and Salt Tolerance of Brassica campestris (rapa)

When plants encounter salinity, growth is reduced initially by water stress and subsequently by toxic levels of ions and by interference with nutrient acquisition and translocation. Calcium (Ca^{2 +}) in particular seems to have an important role in salt tolerance and there are reports of a beneficial effect of increasing Ca^{2 +} availability. Higher potassium (K⁺) concentrations in plants may also improve salinity tolerance as sodium (Na)⁺/K⁺ ratios have been shown to be important. Previous work with a range of Acacia species has suggested that endogenous seed Ca^{2 +} and K⁺ concentrations might influence salinity tolerance at germination, but this has not previously been tested with a single species. The objectives of this investigation were thus to determine whether (1) altered Ca^{2 +} and K⁺ nutrition of Brassica campestris (rapa) L. plants affects the yield and ion content of their seeds, and (2) seeds with different Ca^{2 +} and K⁺ contents differ in their salinity tolerance. Plants were grown in a growth room or greenhouse in (1) Gem® horticultural sand (2) Silvaperl® perlite and sand (2:1), or (3) Shamrock® Medium General Purpose Irish Sphagnum Peat and Vermiperl® vermiculite (1:1). Plants in each growth substrate were supplied with nutrient solutions based on a modified Hoagland's solution as a control, low Ca^{2 +} and low K⁺ solutions containing those elements at half the control strength, but all other mineral elements as far as possible at control strength, and high Ca^{2 +} and high K⁺ solutions containing those elements at double control strength but all other mineral elements, as far as possible, at control strength. An increase in substrate available Ca^{2 +} and K⁺ resulted in increased Ca^{2 +} and K⁺ concentration in seeds, respectively, and was accompanied by a reduction in seed K⁺ and Ca^{2 +}, respectively. The Ca^{2 +} and K⁺ concentrations of seeds affected their salinity tolerance. Increases in seed Ca^{2 +}, K⁺ or Ca^{2 +}+ K⁺ concentrations decreased salinity tolerance at germination. The results, especially in terms of Ca^{2 +} nutrition, contradict previous results of an increased salinity tolerance with increased Ca^{2 +} and/or K⁺ concentrations.     

Long Chain Acyl-Coenzyme A Synthetase 4 (BnLACS4) Gene from Brassica napus Enhances the Yeast Lipid Contents

Long-chain acyl-Coenzyme A (CoA) synthetases (LACSs) catalyze the formation of long-chain acyl-CoA, and play important roles in fatty acid metabolism including phospholipids, triacylglycerol (TAG) biosynthesis and fatty acid β-oxidation. Here, we report the characterization of a LACS gene from Brassica napus. It is highly homologous to Arabidopsis LACS4 and thus designated as BnLACS4. The cloned gene BnLACS4 could complement a LACS-deficient yeast strain YB525. It is mainly expressed in flowers and developing seeds where lipid biosynthesis is at high rate in Brassica napus. When transiently expressed in tobacco leaves, BnLACS4 is localized in endoplasmic reticulum (ER), the common site for eukaryotic pathway of lipid biosynthesis. Expression of BnLACS4 in the yeast strain pep4 increased its lipid content. Taken together, our results suggest that BnLACS4 may be involved in lipid biosynthesis in B. napus.     

大白菜花药培养胚状体诱导研究

以9个大白菜一代杂种为材料进行花药培养。结果表明,大白菜胚状体诱导率受基因型、预处理时间等因素影响。花药在接种后的15～30 d出现胚状体,最后得到了再生植株。     

Transformation of Chinese Cabbage (Brassica rapa L. ssp.pekinensis) by Agrobacterium Micro-Injection into Flower Bud

We obtained two lines of Chinese head cabbage (Brassica rapa L. ssp. pekinensis) selfed progenies containing both an anti-sense gene of BcpLH and a gene for resistance to kanamycin by micro-injecting buds of their primary transformants (T0) with Agrobacterium tumefaciens strain LBA4404. 31 positive plants resistant to kanamycien were recovered. Southern blot analysis confirmed the presence of T-DNA in two transgenic plants. One (DHZ-13-1) exhibits the characteristics of out-toward rosette and cauline leaves, and nested flower model in which secondary complete flower developed from the base of the primary ovary and the third flower from the ovary in the secondary flower, and so on, while another(DHZ-6-1) has no phenotype change. ABA and IAA affected the root growth of progeny of DHZ-13-1, but 6-BA was insensitive to hypocotyl growth during its seedling development.     

2个生理指标与大白菜耐抽薹性相关性初探

分析了白菜不同抽薹特性品种在低温诱导下POD(过氧化物酶)、CAT(过氧化氢酶)活性氧清除系统数值与白菜抽薹特性的关系,为白菜耐抽薹育种及快速选择抽薹特性品种提供理论依据,以20个不同抽薹特性的结球大白菜为材料,评价品种抽薹特性并测定低温诱导后叶片POD和CAT含量。结果表明, POD对照组,冷处理组及冷处理组比对照组与品种的抽薹特性显著正相关,而CAT对照组与抽薹特性负相关,冷处理组及冷处理组比对照组与抽薹性显著正相关。结论:接球大白菜的耐抽薹性与品种在低温胁迫下的活性氧清除能力有一定的相关性。     

大白菜抗软腐病基因BrWRKY33植物表达载体的构建

利用农杆菌侵染法获得抗软腐病转BrWRKY33基因大白菜,首先要构建植物表达载体。本实验根据BrWRKY33基因的序列及pROK2表达载体的酶切位点设计引物(FN/RN),获得BrWRKY33基因,然后将该基因连接到克隆载体pGEM-TEasy,重新构建了克隆载体T-WRKY。经测序及酶切验证,选取PCR反应中错配率最低的产物,构建了表达载体pROK2-WRKY,并将该表达载体转入到根癌农杆菌EHA105中,并对其转化子进行了鉴定。结果表明,PCR扩增获得目的基因BrWRKY33全长约1443bp,PCR及酶切结果鉴定表明克隆载体T-WRKY已经重新构建成功。表达载体的PCR及BamHⅠ和KpnⅠ双酶切鉴定表明有4个重组子表现阳性,表明目的基因已经插入到pROK2表达载体,表达载体构建成功。PCR鉴定表明表达载体pROK2-WRKY已经转入根癌农杆菌EHA105中。本实验结果将为进一步研究大白菜抗软腐病基因的转化奠定基础。     

A Chronic Plant Test for the Assessment of Contaminated Soils. Part 1: Method development (9 pp)

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PREAMBLE According to the German Federal Soil Protection Act (BBodSchG 1998), the habitat function of soils must be protected. Despite the fact that in the Federal Soil Protection Ordinance (BBodSchV 1999) it has not been established how this goal can be reached reliably, it is clear that such a biological function can only adequately be assessed using biological test methods. This is especially true when a soil is contaminated by a mixture of often unknown chemicals. In such a case the use of chemical analysis aiming at a small range of known substances is not sufficient and must therefore be supplemented by biological methods. For this reason, several standardised test methods are available (e.g. using earthworms, collembolans or plants; Römbke and Knacker 2003; ISO 2003). Since acute tests are usually not sensitive enough for the assessment of potentially contaminated soils (e.g. Hund-Rinke et al. 2002), chronic tests like the earthworm reproduction tests (ISO 1998) are recommended for this purpose.

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A chronic plant test for the determination of phytotoxicity was missing until quite recently. The term phytotoxicity is understood here as the capacity of a compound or a contaminated soil to cause temporary or long-lasting damage to plants (EPPO 1997). Therefore, the German Ministry for Education and Research sponsored a project (1997 – 1999) in which – based on existing standardised methods – such a chronic plant laboratory test was developed and partly validated (Kalsch and Römbke 2000). The new test can be used for the evaluation of single chemicals (see Part 1 of this mini-series) as well as for the assessment of contaminated or remediated soils (see Part 2 of this mini-series).

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ABSTRACT Background and Scope. As part of the efforts to improve the biological testing of contaminated soils, the German government sponsored the standardisation of a chronic plant bioassay. This new test is based on experiences with various acute plant tests (e.g. published by OECD or ISO) and existing North American Plant-Life-Cycle Bioassays. In this contribution the characteristic properties of the test are described.

Methods

The test can be performed either with Brassica rapa (turnip) or Avena sativa (oat). Its duration is 35 to 64 days with OECD artificial soil and a German standard field soil acting as controls. Water and nutrients are provided by an automatic wick irrigation system. Besides measuring biomass and shoot length, the number of pods, seeds and flowers are applied as chronic measurement endpoints. During the development of the test, TNT (2,4,6-trinitrotoluene) and Pyrene were used as model test substances.

Results

Pyrene did not affect B. rapa (turnip) in concentrations of up to 10.000 mg/kg soil (due to the often low sensitivity of A. sativa (oat) no further test with this substance was performed). Depending on the endpoint the results varied in the tests with TNT. With few exceptions, the NOEC (No Observed Effect Concentration) values were determined as 55.5 mg TNT/kg soil for B. rapa (turnip) and as 75 (unfertilised) and 150 (fertilised) mg TNT/kg soil for A. sativa (oat). The EC50-values varied between 96.3 and 207.2 mg TNT/kg soil for B. rapa (turnip) and 183.1 – 505.6 mg TNT/kg soil for A. sativa (oat), depending on the endpoint.

Outlook

The results of this work have been used to prepare a draft test guideline, which has recently been standardised by the International Organisation for Standardisation (ISO). Practical experiences with this test system are described in Part II of this mini series.     

Effect of regeneration procedures on genetic diversity in Brassica napus and B. rapa as estimated by isozyme analysis

This investigation was conducted to assess changes in the genetic structure of two varieties of two species of annual Brassica. Seeds of B. napus cv. Topas and B. rapa Broccoletto were sent to nine research institutes in different geographical areas of Europe for regeneration. The multiplied material was sent back after one year of regeneration and analysed electrophoretically. The original populations of each species and their multiplied samples were stained for 12 different enzymes, of which 4 were found to be polymorphic (DIA, SKD, GPI and PER). It was possible to detect considerable differences in isozyme patterns in B. napus and allelic frequencies in B. rapa, both within and between populations. When the original population was compared with the regenerated samples, the Chi-square homogeneity test for all pairwise comparisons revealed distinctness with a 99% probability for B. napus and 95% probability for B. rapa with one or more of the enzyme systems examined. Furthermore, the average of gene diversity analysis (Nei, 1973) revealed that some regenerated populations have less while others have increased genetic variation compared with the original population. These observations indicated that the frequencies were non-random and considerable shifts in genetic diversity have occurred during multiplication. In addition, different regeneration procedures have caused the fixation of certain alleles.     

4种除草剂对胡麻田油菜、荞麦和苦荞麦的防除效果

中国西北和华北地区油菜、荞麦和苦荞麦种植面积较大,其收获时遗落在土壤中的种子翌年出苗后变成了严重危害胡麻的杂草,已成为胡麻生产中亟待解决的突出问题。笔者对2甲·辛酰溴等4种除草剂防除胡麻田油菜、荞麦和苦荞麦开展了应用试验,结果表明：40% 2甲·辛酰溴乳油、25%辛酰溴苯腈乳油和80%溴苯腈可溶粉剂3种除草剂是防除胡麻田油菜的高效除草剂,在600、450、480 g/hm²剂量下药后45天的株防效和鲜重防效均达100%,胡麻增产率分别为83.24%、87.79%、88.13%,3种除草剂在推荐剂量下对胡麻安全,其大面积示范推广的适宜剂量分别为600、450、480 g/hm²。40% 2甲·辛酰溴乳油、25%辛酰溴苯腈乳油、80%溴苯腈可溶粉剂和24%氨氯吡啶酸水剂4种除草剂是防除胡麻田荞麦的高效除草剂,在660、450、540、180 g/hm²剂量下药后45天的株防效在90.32%~98.07%,鲜重防效在97.30%~99.27%,胡麻增产率在87.31%~92.80%,4种除草剂在推荐剂量下对胡麻安全,其大面积示范推广的适宜剂量分别为660、450、540、180 g/hm²,24%氨氯吡啶酸水剂需使用防护罩喷头进行定向喷雾。24%氨氯吡啶酸水剂是防除胡麻田苦荞麦的高效除草剂,在216 g/hm²剂量下药后45天的株防效和鲜重防效分别为91.42%和98.71%,胡麻增产率为94.24%,此种除草剂在使用防护罩喷头进行定向喷雾条件下对胡麻安全,其大面积示范推广的适宜剂量为216 g/hm²。