不同形态微生物菌剂对不结球白菜生长和品质的影响

合理施用微生物菌剂可显著改善蔬菜土壤环境,增加产量,提升品质.本试验以不结球白菜为供试材料,在设施条件下设置不施微生物菌、施用液态和固态微生物菌剂处理,研究不同形态微生物菌剂对不结球白菜生长和品质的影响.结果表明:施用液态和固态微生物菌剂均能显著提高土壤脲酶活性,增加植株全氮含量,增大叶面积,显著提升叶片SPAD值,提...     

秸秆浸提液漂浮栽培对不结球白菜产量及相关品质特性的影响

以1/4园试营养液为对照,以腐熟发酵的农业秸秆(苋菜和番茄植株残体)浸提液模拟富含N、P的水体进行不结球白菜(Brassica campestris ssp.chinensis Makino)的漂浮栽培,对茎叶的单株鲜质量和干质量及叶片的VC、可溶性蛋白质、蔗糖、果糖、葡萄糖含量及可溶性糖总含量进行了比较分析。结果表明:在24 d的栽培周期内,随生长时间的延长,处理组和对照组茎叶的单株鲜质量和干质量均呈增加趋势,但从第10天开始,处理组茎叶的单株鲜质量和干质量均显著低于对照组。栽培初期叶片VC含量均最高且处理组叶片VC含量显著低于对照组;从第10天开始,叶片VC含量降低,但处理组叶片VC含量与对照组无显著差异。处理组与对照组叶片可溶性蛋白质含量均呈波动的变化趋势,但总体差异不显著。处理组叶片蔗糖含量总体上与对照组差异不显著,仅在第24天显著低于对照组。处理组叶片的果糖含量均高于对照组,特别是在第24天为对照组的1.81倍,差异显著。叶片葡萄糖含量均呈先升高后降低的趋势,可溶性糖总含量则呈波动的变化趋势,对照组叶片葡萄糖含量和可溶性糖总含量总体上高于处理组,但差异不显著。研究结果显示:用秸秆浸提液漂浮栽培,虽然不结球白菜的生长量(即产量)有所减少,但相关的品质指标变化不大,对果糖合成和积累还有一定的促进作用,因而,此种秸秆浸提液可用于栽培商品不结球白菜。     

The construction of a genetic linkage map of non-heading Chinese cabbage （Brassica campestris ssp. chinensis Makino）

Non-heading Chinese cabbage （Brassica carnpestris ssp. chinensis Makino） is one of the most important vegetables in eastern China. A genetic linkage map was constructed using 127 doubled haploid （DH） lines, and the DH population was derived from a commercial hybrid ＂Hanxiao＂ （lines SW-13 x L-118）. Out of the 614 polyrnorphic markers, 43.49% were not assigned to any of the linkage groups （LGs）. Chi-square tests showed that 42.67% markers were distorted from expected Mendelian segregation ratios, and the direction of distorted segregation was mainly toward the paternal parent L-118. After sequentially removing the markers that had an interval distance smaller than 1 cM from the upper marker, the overall quality of the linkage map was increased. Two hundred and sixty-eight molecular markers were mapped into 10 LGs, which were anchored to the corresponding chromosome of the B. rapa reference map based on com- mon simple sequence repeat （SSR） markers. The map covers 973.38 cM of the genome and the average interval distance between markers was 3.63 cM. The number of markers on each LG ranged from 18 （R08） to 64 （R07）, with an average interval distance within a single LG from 1.70 cM （R07） to 6.71 cM （R06）. Among these mapped markers, 169 were sequence-related amplified polymorphism （SRAP） molecular markers, 50 were SSR markers and 49 were random amplification polymorphic DNA （RAPD） markers. With further saturation to the LG9 the current map offers a genetic tool for loci analysis for important agronomic traits.     

弱光对不结球白菜光合特性与叶绿体超微结构的影响

研究了不结球白菜耐弱光品种“矮王”和不耐弱光品种“绿优”经弱光处理后,光合特性和叶绿体超微结构的变化.结果表明:遮荫处理后,“矮王”的总叶绿素和叶绿素b含量大幅度上升;2个品种的净光合速率（P_n）、光补偿点（LCP）、光饱和点（(LSP）、表观量子效率（AQY）、羧化效率（CE）均呈下降趋势;除光补偿点外,“矮王”的下降幅度均小于“绿优”.叶绿素荧光参数分析表明,在弱光条件下,“绿优”的光系统Ⅱ（PSII）受到伤害,F_v/F_m、ETR、Φ_PSⅡ和q_P的下降幅度均大于耐弱光品种“矮王”.电镜超微观察结果显示:“绿优”的叶绿体内部出现裂缝和孔洞,多数基质片层断裂,基粒片层解体,线粒体出现内含物减少、发生空洞的现象;而“矮王”的叶绿体结构良好,线粒体发育正常.表明弱光处理可以影响植物的光合机构,但对不同品种的影响不同.     

Effects of molybdenum on ascorbate-glutathione cycle metabolism in Chinese cabbage (Brassica campestris L. ssp. pekinensis)

A hydroponic trial was conducted to investigate effects of molybdenum (Mo) on ascorbate-glutathione cycle (AsA-GSH cycle) metabolism in Chinese cabbage (Brassica campestris L. ssp. pekinensis). Mo was applied at four rates: 0, 0.01, 0.15 and 1.5 mg l⁻¹. The concentrations of ascorbate, dehydroascorbate, reduced- and oxidized- glutathione, and activities of five key enzymes in the AsA-GSH cycle were studied. The results showed that appropriate Mo application increased the fresh weight of Chinese cabbage, but excess application of Mo (1.5 mg l⁻¹ Mo) decreased the fresh weight. Total ascorbate and reduced ascorbate concentrations in the Chinese cabbage increased with Mo application rates. Although no significant differences existed in DHA concentration between the different Mo regimes, but it has an increase trend with the 0.01 mg l^-1 Mo treatment, and then decreased with the Mo level increasing. No significant difference in GSH concentration was found between the different Mo treatments. Compared with the control, the GSSG concentration decreased significantly in the 0.01 mg l⁻¹ Mo treatment. The activities of APX, MDHAR, DHAR and GR increased due to Mo application. But the activity of AAO decreased with increasing Mo application rates. It is hypothesized that Mo may promote the redox process and regeneration of ascorbic acid, and affect the ability of anti-oxidation in the Chinese cabbage. Responsible Editor: Jian Feng Ma.     

不结球白菜叶子重量性状遗传模型分析

应用主基因+多基因6个世代联合分离分析方法, 对不结球白菜SI×秋017组合的叶片重和叶柄重性状进行了分析。结果表明, SI×秋017组合的叶片重性状遗传受1对负向完全显性主基因+加性-显性多基因(D-4)控制, 主基因加性效应为1.8991, 显性效应为-1.8991; 多基因加性效应为-1.2934, 显性效应为1.7933; 势能比值为-1.3865, 显性度为-1.0000; B1、B2和F2世代群体叶片重的主基因遗传率分别为6.98%、4.33% 和36.08%; B1、B2和F2世代群体叶片重的多基因遗传率为16.03%、7.39%和23.96%。叶柄重的遗传受1对加性主基因+加性-显性多基因(D-2)控制, 主基因加性效应为-1.1457, 显性效应为0; 多基因加性效应为1.3472, 多基因显性效应为2.5788; 势能比值为1.9142, 显性度为0。B1、B2和F2世代群体叶柄重的主基因遗传率分别为31.72%、5.27%和57.94%。B1、B2和F2世代群体叶柄重的多基因遗传率分别为0.42%、4.59%和4.80%。对SI×秋017组合叶片重性状的改良要在晚代选择; 对叶柄重的改良要以主基因为主, 可在早代选择。     

春化温度对大白菜花芽分化和抽薹的影响

以冬性有明显差异的 6个大白菜品种为试材 ,研究了 4个春化温度对大白菜的花芽分化和抽薹的影响 .结果表明 ,弱冬性材料在 3～ 11℃范围内 ,花芽分化和抽薹均能在较短的时间完成 ;强冬性材料适宜的春化温度为 3～ 7℃ ,超过 7℃ ,花芽分化和抽薹急剧延迟 ;冬性中等材料适宜的春化温度为 3～ 9℃ .故一般采用 3～ 7℃的春化温度对不同冬性材料均适宜     

Plastid differentiation and chlorophyll biosynthesis in different leaf layers of white cabbage (Brassica oleracea cv. capitata)

The contents of protochlorophyllide, protochlorophyll and chlorophyll together with the native arrangements of the pigments and the plastid ultrastructure were studied in different leaf layers of white cabbage (Brassica oleracea cv. capitata) using absorption, 77 K fluorescence spectroscopy and transmission electron microscopy. The developmental stage of the leaves was determined using the differentiation of the stoma complexes as seen by scanning electron microscopy and light microscopy. The pigment content showed a gradual decrease from the outer leaf layer towards the central leaves. The innermost leaves were in a primordial stage in many aspects; they were large but had typical proplastids with few simple inner membranes, and contained protochlorophyllide and its esters in a 2 : 1 ratio and no chlorophyll. Short‐wavelength, not flash‐photoactive protochlorophyllide and/or protochlorophyll forms emitting at 629 and 636 nm were dominant in the innermost leaves. These leaves also had small amounts of the 644 and 654 nm emitting, flash‐photoactive protochlorophyllide forms. Rarely prolamellar bodies were observed in this layer. The outermost leaves had the usual characteristics of fully developed green leaves. The intermediary layers contained chlorophyll a and chlorophyll b besides the protochlorophyll(ide) pigments and had various intermediary developmental stages. Spectroscopically two types of intermediary leaves could be distinguished: one with only a 680 nm emitting chlorophyll a form and a second with bands at 685, 695 and 730 nm, corresponding to chlorophyll–protein complexes of green leaves. In these leaves, a large variety of chloroplasts were found. The data of this work show that etioplasts, etio‐chloroplasts or chloro‐etioplasts as well as etiolated leaves do exist in the nature and not only under laboratory conditions. The specificity of cabbage leaves compared with those of dark‐grown seedlings is the retained primordial or intermediary developmental stage of leaves in the inner layers for very long (even for a few month) period. This opens new developmental routes leading to formation of specially developed plastids in the various cabbage leaf layers. The study of these plastids provided new information for a better understanding of the plastid differentiation and the greening process .     

Agrobacterium-mediated transformation of cotyledonary explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis)

 A procedure for producing transgenic Chinese cabbage plants by inoculating cotyledonary explants with Agrobacterium tumefaciens strain EHA101 carrying a binary vector pIG121Hm, which contains kanamycin-resistance and hygromycin-resistance genes and the GUS reporter gene, is described. Infection was most effective (highest infection frequency) when explants were infected with Agrobacterium for 15 min and co-cultivated for 3 days in co-cultivation medium at pH 5.2 supplemented with 10 mg/l acetosyringone. Transgenic plants of all three cultivars used were obtained with frequencies of 1.6–2.7% when the explants were regenerated in shoot regeneration medium solidified with 1.6% agar. A histochemical GUS assay and PCR and Southern blot analyses confirmed that transformation had occurred. Genetic analysis of T1 progeny showed that the transgenes were inherited in a Mendelian fashion. Received: 15 December 1998 / Revision received: 2 July 1999 · Accepted: 8 July 1999     

Developmental characteristics of floral organs and pollen of Chinese cabbage (Brassica campestris L. ssp. chinensis)

Morphological and cytological studies are complementary approaches to understand the molecular mechanisms that regulate floral developmental pathways. To better understand abnormal mutant phenotypes in floral development, we conducted detailed observations and investigations of the morphology, cytology, and cell ultrastructure of wild-type Chinese cabbage (Brassica campestris L. ssp. chinensis Makino and syn. B. rapa ssp. chinensis) flowers when they developed from primordia to anthesis. First, we measured bud and organ length with a stereo microscope and observed the developmental status and characteristics of the floral organs using a scanning electron microscope; then we made thin slices of anthers to observe the developmental stage and characteristics of pollen using an optical microscope; and finally, we made super-thin slices of anthers to observe the ultrastructure of pollen during its development with the aid of a transmission electron microscope. In this study, the floral developmental continuum was divided into 17 stages based on significant changes in the shape of floral primordia, and the pollen developmental continuum was divided into 14 stages based on the developmental characteristics. The results could provide the morphological basis for further research on the molecular mechanisms that regulate development of the floral organs and/or pollen of Chinese cabbage and their allied species.     

紫外线-B照射对小白菜生长、产量及品质的影响

在温室条件下,采用辐照度为0.4、0．5和0．6W·m^-2的紫外线-B（UV—B）对小白菜（Brassi cacampestrisssp．chinensis Makino）照射2、3和4h·d^-1,连续照射7d后,研究了UV—B照射对小白菜生长、产量及品质的影响。结果表明,停止照射后35d,各处理组小白菜的鲜质量、株高、叶片数、干物质含量、叶绿素含量、可溶性糖含量及硝酸盐含量均与对照无显著差异,但Vc含量增加,可溶性蛋白质含量降低。经UV—B照射后,下胚轴长度大于2cm的小白菜植株所占的百分率显著低于对照,说明UV—B照射可有效控制小白菜植株徒长。从子叶充分展开时用0,4—0．6W·m^-2 UV—B照射2h·d^-1,连续照射7d,既能有效控制小白菜植株徒长、提高Vc含量,又不会造成产量和品质的下降,为最佳的补充UV—B处理条件。     

Mutation in EMB1923 gene promoter is associated with chlorophyll deficiency in Chinese cabbage (Brassica campestris ssp. pekinensis)

Leaf color mutants are widespread in higher plants and can be used as markers in crop breeding or as important material in understanding the regulatory mechanisms of chlorophyll biosynthesis and chloroplast development. A stably inherited plant etiolated mutation (pem) was obtained from its wild‐type ‘FT’ (a doubled haploid line of the Chinese cabbage variety ‘Fukuda 50’) by combining ⁶⁰Co‐γ radiation and isolated microspore culture in Chinese cabbage. Compared to the wild‐type ‘FT’, the chlorophyll content in the pem mutant was decreased, the photosynthetic capacity was reduced and the chloroplast development was retarded. These physiological changes may lead to a reduction in growth and yield in the pem mutant line. Genetic analysis showed that the mutant phenotype was controlled by the single recessive nuclear pem gene. The pem gene was mapped to a 25.88 kb region on the A03 chromosome. Cloning and sequencing results showed that there was only one DNA sequence variation in this region, which was a 30 bp deletion on the promoter of Bra024218. Its homologous gene encodes EMBRYO DEFECTIVE 1923 (EMB1923) in Arabidopsis thaliana. We therefore predicted that Bra024218 was the mutated gene associated with etiolated leaves in Chinese cabbage. The pem mutant is a useful line for researching chloroplast development and the mechanism of leaf color mutation in Chinese cabbage.     

Management of textile wastewater for improving growth and yield of field mustard (Brassica campestris L.)

Disposal of industrial wastewater is a current issue of urbanization. However, this problem can be sorted out by using wastewater as an alternate source of irrigation after the addition of some amendment. In this way, the problem of disposal of wastewater not only will be resolved but also scarcity of irrigation water can be kept off in the future. The current research study was performed to evaluate the influence of different concentrations of wastewater along with canal water for enhancing growth and yield of field mustard. Plants were irrigated with different mixtures of canal water and wastewater (75:25, 50:50, 25:75, and 00:100) in addition to canal water as control. The results revealed that application of 50:50% waste and canal water improved plant height, the number of pods plant^?1, pod length, root length, root dry weight, shoot dry weight, 100 grain weight, grain and biomass yield plant^?1, and nitrogen, phosphorus, and potassium concentration in grain and straw up to 16%, 15%, 17%, 29%, 15%, 56%, 25%, 41%, 35%, 20%, 52%, 45%, 20%, 44%, and 42%, respectively, over positive control treatment. While, nutrient uptakes and agronomic efficiency of fertilizers also improved by the application of 50:50% canal and wastewater compared to positive control treatment. Furthermore, the concentration of heavy metals, predominantly Cr, Cu, Cd, and Pb, was reduced in grains by application of 50% canal water and 50% wastewater. The outcomes suggest that wastewater utilization along with canal water mixing might be an effective approach for enhancing growth and yield of field mustard.     

Isolated microspore culture of Chinese flowering cabbage (Brassica campestris ssp. parachinensis)

Microspores of several genotypes of Brassica campestris ssp. parachinensis have been cultured in vitro and induced to undergo embryogenesis and plant formation. Conditions favourable for embryogenesis in this species include a bud size of 2–2.9 mm, NLN-13 culture medium (Nitsch and Nitsch 1967; Lichter 1981, 1982; Swanson 1990), and an induction through exposure to 32°C for a period of 48 h. Longer periods of an elevated temperature for induction of embryogenesis resulted in embryo abortion at early developmental stages. With the protocol developed here, microspores of 60–80% of donor plants could be induced to produce embryos, although embryo yields were low, i.e. 2–5 embryos per 10 buds. Some genotypes responded to culture conditions with high numbers of embryo formation (100–150 embryos per 10 buds) but most of these subsequently failed to mature. The pattern of cell division and morphological changes of the microspores in culture were studied using various microscopic techniques.     

Inheritance of seed colour in turnip rape (Brassica campestris L.)

Summary A high gene frequency for ps (parallel spindles) is expected in cultivated tetraploid potatoes, S. tuberosum Group Tuberosum, if 2n pollen produced by ancestral diploid plants which were psps was involved in the origin and evolution of the potato. Fifty-six North American cultivars (varieties and advanced selections) were pollinated by diploid clones, either W 5295.7 or W 5337.3 which are homozygous recessive for ps. The segregation ratios in regard to 2n pollen production in derived tetraploid progenies, from 4x×2x crosses, reveal the genotype of ps in the cultivars. Microsporogenesis of 2n pollen producing 4x progeny was observed to avoid an overestimation of the frequency of 2n pollen producing plants due to mechanisms other than parallel spindles. More than 50% of the 56 cultivars are simplex (Pspspsps), since in each of these cultivars about 50% of their progeny produced 2n pollen. The ps gene frequency in the 56 cultivars was estimated as high as 0.69. The high frequency of ps in the tetraploid cultivars clearly supports the hypothesis that 2n pollen produced by plants homozygous recessive for ps have been involved in the origin of cultivated tetraploid potatoes, since a higher frequency of ps in the tetraploid than in the ancestral diploid population can be expected from sexual polyploidization but not from somatic doubling. The importance of meiotic mutants such as ps for the successful evolution of polysomic polyploids is emphasized.     

不结球白菜根尖体细胞染色体制片及其二倍体和四倍体有丝分裂过程观察

以不结球白菜(Brassica campestris ssp.chinensis Makino)根尖为材料,对染色体制片过程中根长(小于0.5 cm、0.5 ～ 1.0 cm、1.0～2.0 cm及大于2.0 cm)、预处理方法、预处理剂种类(2 mmol·L-1 8-羟基喹啉、2 mmol·L-1 8-羟基喹啉与质量浓度0.2g·L-1秋水仙素等体积混合液、质量浓度0.2g·L-1秋水仙素、饱和对二氯苯、质量浓度20或40 mg·L-1放线菌酮、质量浓度40mg·L-1放线菌酮与2 mmol·L-1 8-羟基喹啉等体积混合液)及预处理时间(1.0～3.5 h)进行了比较和筛选;在此基础上,对二倍体和四倍体不结球白菜根尖体细胞有丝分裂过程进行观察.结果表明:根长度对分裂相的数量有显著影响;根长1.0～2.0 cm,分裂相相对较多,占细胞总数的64.75％.冰冻预处理22 ～ 23 h,能获得一定量的分裂相.采用不同的预处理剂及预处理时间,分裂相的数量及染色体形态有明显差异;用质量浓度40 mg·L-1放线菌酮溶液处理3.5h,分裂相数量最多,但易导致染色体加倍;用质量浓度20mg·L-1放线菌酮预处理3.5h,染色体长且着丝点及随体清晰,且分裂相较多,占细胞总数的53.65％.因而,根长度以1.0 ～2.0 cm为宜,适宜的预处理方法为用质量浓度20 mg·L-1放线菌酮浸泡2.0～3.0 h.二倍体及四倍体不结球白菜根尖体细胞有丝分裂过程基本相似,在有丝分裂的间期、前期、中期、后期和末期染色体的行为基本一致,但在四倍体的有丝分裂过程中会出现多价体、染色体桥、落后染色体、染色体异常分离及内源有丝分裂等异常现象.     

Regulation of nitrate reductase by nitric oxide in Chinese cabbage pakchoi (Brassica chinensis L.)

Nitrate reductase (NR), a committed enzyme in nitrate assimilation, involves generation of nitric oxide (NO) in plants. Here we show that the NR activity was significantly enhanced by the addition of NO donors sodium nitroprusside (SNP) and NONOate (diethylamine NONOate sodium) to the culturing solution, whereas it was decreased by NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (cPTIO). Interestingly, both NO gas and SNP directly enhanced but cPTIO inhibited the NR activities of crude enzyme extracts and purified NR enzyme. The cPTIO terminated the interaction between NR-generated NO and the NR itself. Furthermore, the NR protein content was not affected by the SNP treatment. The investigation of the partial reactions catalysed by purified NR using various electron donors and acceptors indicated that the haem and molybdenum centres in NR were the two sites activated by NO. The results suggest that the activation of NR activity by NO is regulated at the post-translational level, probably via a direct interaction mechanism. Accordingly, the concentration of nitrate both in leaves and roots was decreased after 2 weeks of cultivation with SNP. The present study identifies a new mechanism of NR regulation and nitrate assimilation, which provides important new insights into the complex regulation of N-metabolism in plants.     

SSR and SCAR mapping of a multiple-allele male-sterile gene in Chinese cabbage (Brassica rapa L.)

The genic multiple-allele inherited male-sterile gene Ms in Chinese cabbage (Brassica rapa L.) was identified as a spontaneous mutation. Applying this gene to hybrid seed production, several B. rapa cultivars have been successfully bred in China. A BC₁ population (244 plants) was constructed for mapping the Ms gene. Screening 268 simple sequence repeat (SSR) markers which cover the entire genome of Chinese cabbage was performed with bulked segregant analysis (BSA). On the basis of linkage analysis, the Ms gene was located on linkage group R07. In addition, through the amplified fragment length polymorphism (AFLP) and the sequence-characterized amplified region (SCAR) techniques combining BSA, two SCAR markers which were converted from corresponding AFLP markers flanked the Ms gene. Finally, a genetic map of the Ms gene was constructed covering a total interval of 9.0 cM. Two SCAR markers, syau_scr01 and syau_scr04, flanked the Ms gene at distances of 0.8 and 2.5 cM, respectively. All the SSR markers (cnu_m273, cnu_m030, cnu_m295, and syau_m13) were mapped on the same side of the gene as syau_scr04, the nearest one of which, syau_m13, was mapped at a distance of 3.3 cM. These SSR and SCAR markers may be useful in marker-assisted selection and map-based cloning. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.