十字花科蔬菜基因组含量的测定与分析

以我国14种重要十字花科蔬菜为材料,利用流式细胞术测定基因组合量.其中青苤蓝、乌塌菜、雪里蕻、芥蓝的基因组含量属首次报道.本试验数据与国外文献提供的相关数据对比,发现青萝卜、结球甘蓝、青花菜和根用芥菜的基因组含量与已报道数值基本吻合,而大白菜、花椰菜的基因组含量值与报道数据存在差异.造成同一物种基因组含量值差异的因为可能是品种的不同,也可能与生长环境或测定时参考标准选用等因素不同有关.     

芥蓝与芸苔属几个常见种花粉形态的比较

本文通过对芸苔属(Brassica L.)几个常见种芥蓝(B. alboglabra Bailey)、芥菜(B.juncea(L. )Coss.)、油菜(B. campestris L.)和花椰菜(B. oleracea var. botrytis L.)等花粉形态的光学显微镜和扫描电镜的观察比较,得知它们的花粉形态特征比较一致。而与杨萍等关于芥蓝的花粉形态特征——无极,具散孔这一报道截然不同;亦与黄增泉(Huang Tseng-chieng)关于芥菜的花粉特征——不具萌发孔的报道不同。作者的研究表明,上述几个种的花粉均具3沟,沟长而明显。外壁具网状纹饰,网眼较粗。但种与种间其花粉大小有所不同。作者不同意杨萍等关于“由于芥蓝的花粉形态与芸苔属其它种的花粉明显区别,说明芥蓝与芸苔属其它种不很亲缘”的观点,而认为,从花粉形态看,芥蓝与芸苔属其它种是基本一致的,把它们置于同一自然类群是合理的。     

Comparison of feeding behaviour of two Brassica pests Brevicoryne brassicae and Myzus persicae on wild and cultivated brassica species

Feeding behaviour of the specialist Brassicae aphid, Brevicoryne brassicae (L) (cabbage aphid) and the generalist, Myzus persicae, (Sulzer) (peach potato aphid) was monitored electronically on the susceptible cauliflower, Brassica oleracea var. botrytis cv Newton Seale, and a range of 17 Brassica species, B. carinata, B. juncea, B. nigra, B. macrocarpa, and B. villosa var. drepanensis and cultivated brassica varieties, B. oleracea, B. campestris and B. napus. Aphids, monitored for 10 h on the underside of leaves, performed recognisable feeding behaviour on all brassica species. The main differences in feeding behaviour, between M. persicae and B. brassicae, on the susceptible cauliflower Newton Seale, were fewer probes, shorter times to initially reach the phloem but longer times to establish sustained phloem ingestion and the longer times spent, by M. persicae, in xylem ingestion.Feeding behaviour on the range of brassica species tested indicated that generalist and specialist aphids are influenced differently by the host plant. A longer time spent in xylem ingestion was again the major difference in the feeding behaviour of the two aphids. In addition, rejection of passive phloem ingestion, by M. persicae, was not related so closely to increased time spent in non probing activities, as for B. brassicae. This observation indicates that M. persicae does not generally accept or reject brassica species due to the presence of phagostimulants, such as glucosinolates at the leaf surface or along the stylet pathway, unless the concentration is very high. Differences in feeding strategies employed by generalist and specialist aphids on the same plants are discussed.     

The self-incompatibility (S) haplotypes of Brassica contain highly divergent and rearranged sequences of ancient origin.

In Brassica, the recognition of self-related pollen by the stigma is controlled by the highly polymorphic S locus that encodes several linked and coadapted genes and can span several hundred kilobases. We used pulsed-field gel electrophoresis to analyze the structure of different S haplotypes. We show that the S2 and S13 haplotypes of Brassica oleracea contain extensive sequence divergence and rearrangement relative to each other. In contrast, haplotypic configuration is more conserved between B. oleracea S13 and B. campestris S8, two haplotypes that have been proposed to be derived from a common ancestral haplotype based on sequence comparisons. These results support the view that extensive restructuring of the S locus preceded speciation in Brassica. This structural heteromorphism, together with haplotype-specific sequences, may suppress recombination within the S locus complex, potentially providing a mechanism for maintaining the linkage of coadapted allelic combinations of genes over time.     

传粉导致的转基因白菜与其近缘种属材料间的基因流动

从形态、染色体及分子水平上证实,转基因不结球白菜(Brassica campestris ssp．chinensis var．utilis Tsen et Lee)中编码除草剂Basta抗性的bar基因能在田间条件下,经自然传粉,以较高频率侵入芜菁(B．campestris ssp．rapifera)、结球白菜(B．campestris／sssp．pek／nens／s)和不结球白菜(B．campestrs ssp．ch／nens／s)的基因组中,也能少量侵入同属异种的甘蓝型油菜(B.napus)基因组中;在温室人工辅助授粉条件下,除在上述种中的基因漂移率提高外,bar基因尚能以一定频率侵入同属的黑芥(B．nigra)、埃塞俄比亚芥(B．car／nata)、芥菜(B．juncea)基因组中,但始终未能得到转基因白菜与结球甘蓝(B．oleracea)、萝卜(尺．sativus)的杂种。转基因白菜与十字花科的7种常见杂草经温室人工辅助授粉,也均未得到抗性杂种。     

传粉导致的转基因白菜与其近缘种属材料间的基因流动

从形态、染色体及分子水平上证实,转基因不结球白菜(Brassica campestris ssp. chinensis var. utilis Tsen et Lee)中编码除草剂Basta抗性的bar基因能在田间条件下,经自然传粉,以较高频率侵入芜菁(B. campestris ssp. rapifera)、结球白菜(B. campestris ssp. pekinensis)和不结球白菜(B. campestris ssp. chinensis)的基因组中,也能少量侵入同属异种的甘蓝型油菜(B. napus)基因组中; 在温室人工辅助授粉条件下,除在上述种中的基因漂移率提高外,bar基因尚能以一定频率侵入同属的黑芥(B. nigra)、埃塞俄比亚芥(B. carinata)、芥菜(B. juncea)基因组中,但始终未能得到转基因白菜与结球甘蓝(B. oleracea)、萝卜(R. sativus)的杂种.转基因白菜与十字花科的7种常见杂草经温室人工辅助授粉,也均未得到抗性杂种.     

诸葛菜与芸苔属属间杂交时花粉—雌蕊相互作用的研究

本文对诸葛菜和芸苔属属间杂交的花粉粒萌发和花粉管生长情况进行了研究。其中用诸葛菜作父本,分别和甘蓝型油菜（“文油６号”、“云油３１号”）、“羽衣甘蓝”和白菜型油菜（“凤仪油菜”）杂交时,受精前有一定的亲和性;而和白菜型油菜（“玉溪周永达油菜”）、芥菜型油菜（“泸州四棱”、“南充油菜”）、埃塞俄比亚芥杂交时,存在受精前障碍。同时还发现通过蕾期授粉是克服属间杂交不亲和性的有效方法之一。     

Identification of individual chromosomes and parental genomes in Brassica juncea using GISH and FISH

The three diploid (B. nigra, B. oleracea, B. campestris) and three allotetraploid (B. carinata, B. juncea, B. napus) species of Brassica, known as the "U-triangle" are one of the best model systems for the study of polyploidy. Numerous molecular investigations have provided a wealth of new insights into the polyploid origin and changes during the evolution of Brassica, but there are still many controversial aspects of their relationship and evolution. Interpretation of genome changes during evolution requires individual chromosome identification within the genome and clear distinction of genomes within the allotetraploid. The aim of this study was to identify individual chromosomes of B. juncea (genome AABB; 2n = 4x = 36) and to determine their genomic origin. Fluorescence in situ hybridization with 5S and 45S rDNA probes enabled discrimination of a substantial number of chromosomes, providing chromosomal landmarks for 20 out of 36 chromosomes of B. juncea. Additionally, along with double target genomic in situ hybridization, it allowed assignment of all chromosomes to either the A or B genomes.     

Relationships between growth and gas exchange characteristics in some salt-tolerant amphidiploid Brassica species in relation to their diploid parents

Relationships between growth and different gas exchange characteristics of two amphidiploid salt tolerant species, Brassica napus, and B. carinata with respect to their salt sensitive parents, B. oleracea, and B. nigra were investigated. Twenty three-day old plants of these four species along with those of another amphidiploid moderately salt tolerant B. juncea (developed by hybridization of diploids, B. campestris and B. nigra), and a diploid moderately salt tolerant, B. campestris, were subjected for 28 days to salinized sand culture containing 0, 100 or 200 mol NaCl m(-3) in Hoagland's nutrient solution. The species B. napus and B. carinata produced significantly greater shoot fresh and dry matters than their parents under saline conditions. A close association was found between growth, and assimilation rate for all species differing in degree of salt tolerance. Stomatal conductance (g(s)) was reduced due to salt stress in all species but this variable had no significant correlation with assimilation rate (A). However, the amphidiploid salt tolerant species, B. napus and B. carinata had significantly greater photosynthetic rate, water use efficiency (A/E), intrinsic water use efficiency (A/g(s)) than those of their diploid parents. In conclusion, high salt tolerance of the two amphidiploid species, B. napus and B. carinata was associated with a high assimilation rate, water use efficiency and intrinsic water use efficiency but there was little association of the tolerance of these species with stomatal conductance, leaf water potential or transpiration rate (E).     

Nucleolar dominance does not occur in root tip cells of allotetraploid Brassica species.

Using in situ hybridization and silver staining methods, the numbers of active and inactive rDNA loci have been established for three allotetraploid species of Brassica (B. napus, B. carinata, and B. juncea) and their diploid ancestors (B. campestris, B. nigra, and B. oleracea). The allotetraploid species have chromosome numbers equal to the sum of the numbers in their diploid relatives, but have fewer rDNA loci. All species investigated have lower numbers of active NORs (AgNORs, nucleolar organizer regions) compared with the numbers of rDNA sites revealed by in situ hybridization. The number of active rDNA loci of the allotetraploid species is equal to the number of AgNORs in their diploid ancestors, indicating the absence of nucleolar dominance in amphidiploid Brassica species, at least in root meristematic cells.     

Role of Ca2+ on Growth of Brassica campestris L. and B.juncea (L.) Czern & Coss under Na+ Stress

Root and shoot growth of Brassica campestris L.and B.juncea increased significantly(P0.01) with enhanced Ca2+ treatment along with 60 mM NaCl in the root medium.The maximum fresh mass of shoot and root in B.juncea was recorded at 10 mM Ca2+ concentration.The relative growth rate of shoot of both species reached its maximum at 8 mM of Ca2+ concentration.Average rate of Ca2+ intake(Ca) was higher in B.juncea than B.campestris.In B.juncea,the average transport of Ca2+ to shoot increased by 19%,38%,119%,125% and 169% compared with the control.Furthermore specific utilization rate of Ca2+ was higher in B.juncea than B.campestris.In B.campestris it increased by 9%,32%,41% and 59% at 4,6,8,and 10 mM of calcium in comparison to 2 mM Ca2+ treatment.At 4,6,8 and 10 mM of Ca2+ application,the increase in the leaf area ratio was 10,17,23 and 30%,respectively.In the shoot and root portions of B.campestris and B.juncea,Ca2+ had a linear relationship with potassium and sulfur,whereas it was in antagonism with sodium ion.     

Comparative proteome analysis of Xanthomonas campestris pv. campestris in the interaction with the susceptible and the resistant cultivars of Brassica oleracea

Black rot of cruciferous plants, caused by Xanthomonas campestris pv. campestris , causes severe losses in agriculture around the world. This disease affects several cultures, including cabbage and broccoli, among others. Proteome studies of this bacterium have been reported; however, most of them were performed using the bacterium grown under culture media conditions. Recently, we have analyzed the proteome of X. campestris pv. campestris during the interaction with the susceptible cultivar of Brassica oleracea and several proteins were identified. The objective of the present study was to analyze the expressed proteins of X. campestris pv. campestris during the interaction with the resistant cultivar of B. oleracea . The bacterium was infiltrated in the leaves of the resistant plant and recovered for protein extraction and two-dimensional electrophoresis. The protein profile was compared with that of the bacterium isolated from the susceptible host and the results obtained revealed a group of proteins exclusive to the resistant interaction. Among the proteins identified in this study were plant and bacterium proteins, some of which were exclusively expressed during the resistant interaction.     

Isolation and polypeptide composition of l,3-ß-glucan synthase from plasma membranes of Brassica oleracea

The l,3-ß-glucan synthase (callose synthase, EC 2.4.1.34) was solubilized from cauliflower ( Brassica oleracea L.) plasma membranes with digitonin, and partially purified by ion exchange chromatography and gel filtration [fast protein liquid chromatography (FPLC)] using 3-[(cholamidopropyl)dimethylammonio]-1-propane-sulfonate (CHAPS) in the elution buffers. These initial steps were necessary to obtain specific precipitation of the enzyme during product entrapment, the final purification step. Five polypeptides of 32, 35, 57, 65 and 66 kDa were highly enriched in the final preparation and are thus likely components of the callose synthase complex. The purified enzyme was activated by Ca²⁺, spermine and cellobiose in the same way as the enzyme in situ, indicating that no essential subunits were missing. The polyglucan produced by the purified enzyme contained mainly 1,3-linked glucose.     

Free amino acid content of phloem sap from Brassica plants in relation to performance of Lipaphis erysimi (Hemiptera: Aphididae)

Cultivars of Brassica juncea, B. napus and B. campestris, differing in host-suitability to the mustard aphid (Lipaphis erysimi pseudobrassicae Kalt.), were analysed for their phloem sap content of free amino acids. Sap was collected from excised mustard aphid stylets and analysed by means of high performance liquid chromatography. Aspartic and glutamic acid were, together with glutamine, the dominating amino acids. All plants had very little glycine, γ-aminobutyric acid (GABA), methionine and tryptophan in their sap. This amino acid pattern closely resembled that of cereals. The total concentration and the individual composition of amino acids were not found to be related to the level of aphid resistance previously found in the investigated Brassica species. However, one B. campestris cultivar, yellow sarson YSB-9, on which the aphids were shown to grow more slowly, had less amino acid content than other B. campestris varieties. The slow population growth of aphids on B. juncea, compared to that on B. campestris, is probably due to other than nutritional factors.     

In vivo proteome analysis of Xanthomonas campestris pv. campestris in the interaction with the host plant Brassica oleracea

The genus Xanthomonas is composed of several species that cause severe crop losses around the world. In Latin America, one of the most relevant species is Xanthomonas campestris pv. campestris, which is responsible for black rot in cruciferous plants. This pathogen causes yield losses in several cultures, including cabbage, cauliflower and broccoli. Although the complete structural genome of X. campestris pv. campestris has been elucidated, little is known about the protein expression of this pathogen in close interaction with the host plant. Recently, a method for in vivo analysis of Xanthomonas axonopodis pv. citri was developed. In the present study, this technique was employed for the characterization of the protein expression of X. campestris pv. campestris in close interaction with the host plant Brassica oleracea. The bacterium was infiltrated into leaves of the susceptible cultivar and later recovered for proteome analysis. Recovered cells were used for protein extraction and separated by two-dimensional electrophoresis. Proteins were analysed by peptide mass fingerprinting or de novo sequencing and identified by searches in public databases. The approach used in this study may be extremely useful in further analyses in order to develop novel strategies to control this important plant pathogen.     

Prey-predator relationship between Lipaphis erysimi Kalt. (Hom., Aphididae) and Coccinella septempunctata L. (Col., Coccinellidae). II. Effect of host plants on the functional response of the predator

The present paper examines the effect of the host plants Brassica oleracea, Raphanus sativus and Brassica campestris on which Lipaphis erysimi was reared, on the functional response of the grub of Coccinella septempunctata . The functional response of the grub of C. septempunctata is of type II. The amount of prey consumption increases significantly with the increase of prey density and is maximum in B. oleracea- reared aphids, followed by those reared on R. sativus and B. campestris . The percentage prey consumption declined throughout. This decrease was a maximum for aphids reared on B. campestris followed by R. sativus - and B. oleracea -reared aphids. Analysis of variance also confirmed that the number and percentage prey consumption was significantly influenced between the three host plants and between 12 prey densities. Results show that C. septempunctata has a greater liking for aphids reared on B. oleracea rather than those reared on the other plants that were tested.     

小菜蛾成虫对沙芥的产卵嗜好性的初步研究

通过室内试验,比较了小菜蛾Plutella xylostellaL.对沙生十字花科蔬菜沙芥叶(PugioniumcornutumL.)、小白菜、油菜和甘蓝叶的产卵选择性。结果表明,产卵选择性顺序为沙芥≥小白菜>油菜>甘蓝,在沙芥叶上的产卵率是甘蓝叶上的9倍多;补充营养使小菜蛾成虫的产卵量平均提高2.5倍,产卵次数增加2倍,在有沙芥叶条件下,当日羽化的小菜蛾便可交尾产卵;在无沙芥叶条件下,小菜蛾成虫的产卵前期平均延迟1d左右。     

Reproductive rate of the Indian mustard aphid (Lipaphis erysimi pseudobrassicae) on different Brassica oilseeds: colmparisons with Swedish strains of mustard (Lipaphis erysimi erysimf) and cabbage aphid (Brevicoryne brassicae)

The reproductive rate of the Indian mustard aphid (Lipaphis erysimi pseudobrassicae), a serious pest on Brassica oilseeds in India, was compared with that of Swedish strains of mustard aphid (L. e. erysimi) and cabbage aphid (Brevicoryne brassicae) on six common Indian and Swedish Brassica oilseed cultivars. There were clear differences between L. e. erysimi and L. e. pseudobrassicae in the relative suitability of the cultivars, emphasisipg the biological differences between these aphids. B. brassicae had similar reproductive rates on all cultivars.
In addition, the reproductive rate of L. e. pseudobrassicae was measured on both young and old plants of the six Brassica oilseed cultivars and 14 Indian rai (B. juncea) cultivars. Among the six Brassica cultivars, the reproductive rates were highest on turnip rape and toria (B. campestris), and lowest on yellow sarson (B. campestris) and rai (B. juncea). The ranking of the cultivars was similar for young and old plants. There were no significant differences between the rai (B. juncea) cultivars tested, neither among young nor among old plants. These results from laboratory studies are compared with field data from India on the relative susceptibility of different Brassica oilseed cultivars to L. e. pseudobrassicae.     

烟粉虱对四种蔬菜寄主的选择性

为探讨利用趋避作用防治烟粉虱的可能性,进行了3方面试验：1）烟粉虱Bemisia tabaci (Gennadius) 对黄瓜、花椰菜、油菜和莴苣等4种蔬菜的寄主选择性; 2）互喷烟粉虱不同嗜好的寄主植物汁液对其寄主选择性的影响; 3）不同间作方式对黄瓜叶上烟粉虱成虫密度的影响。结果表明,烟粉虱对供试植物有明显的选择性,其嗜好程度为花椰菜>黄瓜>油菜>莴苣。喷施莴苣和花椰菜汁液对烟粉虱的寄主选择性均有显著影响,喷施莴苣原液后,花椰菜和黄瓜上的成虫数分别减少79.6%和87.4%,花椰菜上的着卵量减少84.3%;喷施花椰菜原液后,莴苣上的成虫数和着卵量分别增加82.4%和79.2%,表明不同蔬菜中的内含物是引起烟粉虱对寄主选择性的重要原因。在黄瓜中间作莴苣可使黄瓜叶上烟粉虱成虫数平均减少76.5%;间作花椰菜时,通过诱集作用,也可使黄瓜叶上的成虫数平均降低69.7%。这些结果为利用喷施蔬菜汁液和间作方式控制烟粉虱危害提供了理论依据。     

Purification and characterization of alpha-amylase from rat pancreatic acinar carcinoma. Comparison with pancreatic alpha-amylase.

alpha-Amylase was purified to apparent homogeneity from normal pancreas and a transplantable pancreatic acinar carcinoma of the rat by affinity chromatography on alpha-glucohydrolase inhibitor (alpha-GHI) bound to aminohexyl-Sepharose 4B. Recovery was 95-100% for both pancreas and tumour alpha-amylases. They were monomeric proteins, with Mr approx. 54000 on SDS/polyacrylamide-gel electrophoresis. Isoelectric focusing of both normal and tumour alpha-amylases resolved each into two major isoenzymes, with pI 8.3 and 8.7. Tumour-derived alpha-amylase contained two additional minor isoenzymes, with pI 7.6 and 6.95 respectively. All four tumour isoenzymes demonstrated amylolytic activity when isoelectric-focused gels were treated with starch and stained with iodine. Two-dimensional electrophoresis, on SDS/10-20%-polyacrylamide-gradient gels after isoelectric focusing, separated each major isoenzyme into doublets of similar Mr values. Pancreatic and tumour-derived alpha-amylases had similar Km and Ki (alpha-GHI) values, but the specific activity of the tumour alpha-amylase was approximately two-thirds that of the normal alpha-amylase. Although amino acid analysis and peptide mapping with the use of CNBr, N-chlorosuccinimide or Staphylococcus aureus V8 proteinase gave comparable profiles for the two alpha-amylases, tryptic-digest fingerprint patterns were different. Antibodies raised against the purified pancreatic alpha-amylase and tumour alpha-amylase respectively showed only one positive band on immunoblotting after gel electrophoresis of crude extracts of rat pancreas and carcinoma, at the same position as that of the purified enzyme. More than 95% of the alpha-amylase activity in the pancreas and in the tumour was absorbed by an excess amount of either antibody, indicating that normal and tumour alpha-amylases are immunologically identical. The presence of additional isoenzymes in the carcinoma, and dissimilarity of tryptic-digest patterns, may reflect an alteration in gene expression or in the post-translational modification of this protein in this heterogeneously differentiated transplantable pancreatic acinar carcinoma.