Distribution of repetitive DNA sequences in Vicia faba chromosomes

In situ hybridization of labelled complementary RNA transcribed from whole DNA to metaphase chromosomes indicates the presence of repetitive DNA in both euchromatin and heterochromatin of the Vicia faba genome.     

Gene organization of chloroplast DNA from the broad bean Vicia faba

Summary Six protein genes have been mapped on broad bean chloroplast DNA by Southern hybridization using the tobacco chloroplast genes as probes. In broad bean chloroplast DNA, the genes for the and subunits of proton-translocating ATPase and the 32,000 dalton thylakoid membrane protein are located near the large subunit gene of ribulose-1,5-bisphosphate carboxylase/oxygenase. The gene for the subunit of proton-translocating ATPase is distantly located from the and genes. The gene for the ribosomal protein CS19 was found close to the ribosomal RNA operon. The gene organization of broad bean chloroplast DNA is therefore quite different from that of tobacco chloroplast DNA. The nucleotide sequence of the spacer region between the large subunit and the genes of the broad bean has been determined. Conserved sequences are found among the putative promoter regions of the chloroplast protein genes.     

Further studies on the mechanism involved in differential staining of BUdR-substituted Vicia faba chromosomes

In a preceding publication we reported that photolysis of BUdR-substituted Vicia faba chromatids occurs during observation with a fluorescence microscope when chromosomes were mounted in a solution containing trypsin and a photosensitive dye (Hoechst 33258 or acridine orange). The present investigations support the hypothesis that the rapid dissolving of the double BUdR-substituted (BB) chromatids observed with our method is due to single-strand breaks induced by a photosensitive dye-visible light system. The agents cysteamine and potassium iodide which reduce BUdR radicals and in this way may inhibit single-strand breaks modify the rate of chromosomes showing differential staining. It was totally suppressed by high cysteamine concentrations and markedly reduced by potassium iodide. Several acridine dyes were tested concerning their ability to induce differential staining. Some of them, e.g. aurophosphine and coriphosphine O, yield good results, others, e.g. acriflavine and acridine yellow, give poor differential staining. In an experiment in which the trypsin concentration was varied to induce approximately optimum and non-optimum digestion conditions the necessity of trypsin treatment in our method was confirmed.     

利用蚕豆根尖细胞微核技术检测Cu、As污染的诱变性

应用蚕豆根尖细胞微核技术检测Cu、As及其复合污染的诱变性能,计数了蚕豆根尖细胞微核千分率(MCN‰)和污染指数(PI)并进行了F检验.结果表明,在一定浓度范围内(Cu＜200mg·L,As＜15mg·L^-1),随着Cu、As浓度的增加,MCN‰上升;24个试验处理组PI均在2以上,各处理组MCN‰有显著差异,和对照组有极显著差异(a＜0,01),表明Cu、As及其复合处理对蚕豆根尖细胞的分裂活动有明显影响.     

Length variation in the non-transcribed spacer of Calliphora erythrocephala ribosomal DNA is due to a 350 base-pair repeat

The non-transcribed spacer regions in the ribosomal DNA cistrons of Calliphora erythrocephala vary in length. This length variation is shown to be due to variable numbers of small repeated units found in certain areas of the NTS² regions, as has been found in several other systems. In contrast to the other organisms, however, the length variation in C. erythrocephala leads to the formation of only two major size classes with a length difference of about 1 kb. We have investigated the nature of this length difference by means of electron microscope heteroduplex and restriction enzyme digestion analyses. We demonstrate that the length variation in C. erythrocephala is due to a variation in the number of 350 bp repeating units defined by the presence of sites for three restriction enzymes, HhaI, Sau3A and AluI. Furthermore, the existence of an XbaI site within one 350 bp unit and the lack of a HhaI restriction site within another 350 bp unit reveals that at least some sequence differences exist among the repeating units. These restriction site differences can be viewed as genetic markers and lead to the hypothesis that the longer NTS class originated from the shorter NTS class by an unequal crossover event that served to duplicate the region containing repetitive units. An interesting feature of our observation is that not all NTS length classes are equally represented, suggesting special rules for the unequal recombination events often hypothesized as the basis for such variation.     

土荆芥挥发性化感物质对蚕豆叶表皮保卫细胞的影响

化感作用是外来植物土荆芥( Chenopodium ambrosioides)成功入侵的机制之一。为了探讨土荆芥挥发油的化感作用机制,该文以蚕豆( Vicia faba)叶的下表皮为材料,将表皮条孵育在分别含土荆芥挥发油、α-萜品烯和对伞花素的MES [2-( N-morpholino) ethanesulfonic acid]缓冲液中,25℃下光照培养30 min,采用吖啶橙/溴乙锭( AO/EB)双荧光染色法和Feulgen染色法,研究土荆芥挥发油、α-萜品烯和对伞花素对保卫细胞活性和细胞核形态的影响。结果表明：在土荆芥挥发油、α-萜品烯和对伞花素的作用下,蚕豆气孔保卫细胞活性降低,细胞核出现固缩、畸形或降解等细胞凋亡特征。随着处理剂量增加,保卫细胞活性显著下降,核异常率显著增加,表明土荆芥挥发油、α-萜品烯和对伞花素均对蚕豆保卫细胞具有细胞毒性,其中,挥发油毒性最大,α-萜品烯的毒性次之,对伞花素的毒性最小;Caspase抑制剂Z-VAD-FMK可缓解挥发油、α-萜品烯和对伞花素对保卫细胞的毒性,提高细胞活性,这种缓解效应随着抑制剂浓度的增加而增大。由此可见,土荆芥挥发油、α-萜品烯和对伞花素诱导蚕豆保卫细胞发生了Caspase依赖性的细胞凋亡。     

The irreversible photoinhibition of the photosystem II complex in leaves of Vicia faba under strong light

Exposure of the photosynthetic machinery to strong light causes the photoinhibition of the photosystem II complex. The recovery from the photoinhibition in vivo was characterized by monitoring the ratio of variable to maximum fluorescence (F_v/F_m) in detached leaves of broad bean (Vicia faba). The changes in the ratio were explained in terms of three components, namely, two saturating exponential components with half rise-times of about 15 and 120 min, respectively, and a non-recovery component. The non-recovery component increased gradually as the exposure to strong light was prolonged. Our results suggest that this irreversible component of the photoinhibition of the photosystem II complex was caused by severe stress due to strong light under which repair of the photosystem II complex was insufficient to allow full recovery. The irreversible photoinhibition is discussed in terms of both the physiology and ecology of plants.     

铅持续胁迫下不同世代蚕豆生理生化指标的变化

为了解长期持续铅胁迫下植物不同世代间生理生化指标的变化,分析植物对铅污染的适应机制及植物对长期持续污染的响应机理,该研究以在250 mg·kg~(-1)Pb持续污染的实验田中种植所得的第4、第8、第10、第13和第15世代蚕豆种子为材料,通过盆栽法,与同世代未受Pb胁迫的蚕豆做对照,分析了与抗氧化酶系统、总代谢等相关的8个生理指标,即叶绿素、蛋白质、可溶性糖、脯氨酸(PRO)、丙二醛(MDA)含量及超氧化物歧化酶(SOD)、谷胱甘肽还原酶(GR)和苹果酸脱氢酶(MDH)活力。结果表明:(1)在长期持续Pb胁迫下,与蚕豆光合作用能力有关的叶绿素含量在第8代与第13代之间与对照组相比具有显著的差异性(P0.05)。(2)可溶性糖含量随胁迫时间的延长而逐渐升高,并在第10代时达到最高,随后开始向无污染的正常水平恢复并趋于稳定;蛋白质含量在胁迫至第13代后开始增加。(3)MDA与PRO含量在胁迫至第10代时达到最高,并随胁迫时间的延长而降低,至第13代后逐渐趋于稳定。(4)抗氧化酶系统中的SOD、GR和MDH也同样呈现出先高后低的趋势,且GR的变化程度最大,MDH活力最早表现出与对照组的最大差异。研究认为:在长期的持续Pb污染胁迫下,蚕豆种群对Pb污染胁迫的适应性不断提高。     

Assessment of the genotoxicity of olive mill waste water (OMWW) with the Vicia faba micronucleus test

The present study concerns the genotoxicity of olive mill waste water (OMWW) generated in mills producing olive oil in Morocco. The Vicia faba micronucleus test was used to evaluate the genotoxicity of OMWW and the six major phenolic compounds identified by HPLC in this effluent. Five dilutions of OMWW were tested: 0.1, 1, 5, 10 and 20%. Maleic hydrazide was used as a positive control. The results showed that OMWW was genotoxic at 10% dilution. In order to investigate the components involved in this genotoxicity, the six major phenols present in this effluent, oleuropein, gallic acid, 4-hydroxyphenyl acetic acid, caffeic acid, paracoumaric acid and veratric acid, were studied at concentrations corresponding to the genotoxic concentration of the OMWW itself. Two phenols, gallic acid and oleuropein induced a significant increase in micronucleus frequency in Vicia faba; the four other phenols had no significant genotoxic effect. These results suggest that under the experimental conditions of our assay, OMWW genotoxicity was associated with gallic acid and oleuropein.     

Relation between growth pattern and resistance to Aphis fabae Scopoli in three varieties of field bean (Vicia faba L.)

Three varieties of Field (Spring) Bean were compared in a small plot trial: Rastatt (known to be relatively resistant to Aphis fabae), Schlanstedt (very susceptible) and an English Tick Bean. Records were kept of the growth of the plants and of the day-to-day incidence of winged Aphis fabae and Acyrthosiphon pisum, with periodic aphicidal spraying to minimize damage to the plants.Schlanstedt was the first to flower and fruit, Rastatt intermediate and Tick last with the most shoots, leaves and flowers. Yields were about the same from Schlanstedt and Rastatt and somewhat lower from Tick. The relative incidence of A. fabae both among the organs of one variety and between varieties was correlated with growth pattern. The smaller number of aphids on Rastatt compared with both Schlanstedt and Tick was associated with the compromise growth pattern of Rastatt, wherein growth was more equally divided between shoots and fruits, and apparently less vigorous in the individual organs, during the critical weeks.These observations, together with the contrasted behaviour of A. pisum and previous work on phloem sap composition in relation to plant growth and aphid nutrition, suggest that the sap may have been least nutritious for aphids in Rastatt, and that the resistance of Rastatt to A. fabae may have been due to the winged migrants responding to some superficial plant features associated with growth pattern and sap composition. Possible practical implications are briefly mentioned.

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Zusammenfassung In einem kleinen Parzellenversuch wurden drei Ackerbohnensorten verglichen: Rastatter (bekannt als relativ resistent gegen Aphis fabae), Schlanstedter (sehr anfällig) und eine englische Tick Bohne. Das Wachstum der Pflanzen und der tägliche Befall mit geflügelten Aphis fabae und Acyrthosiphon pisum wurden aufgezeichnet (protokolliert), dabei hielten periodische Spritzungen mit Aphidiciden die Schädigung der Pflanzen möglichst niedrig.Schlanstedter blühte und fruchtete als erste, danach Rastatter (intermediär) und Tick mit den meisten Trieben, Blättern und Blüten zuletzt. Die Erträge von Schlanstedter und Rastatter waren annähernd die gleichen, die von Tick etwas niedriger. Der relative Befall mit Aphis fabae stand sowohl auf den Organen einer Sorte wie zwischen den Sorten in Beziehung zur Wuchsform. Die geringere Anzahl der Blattläuse auf Rastatter im Vergleich sowohl mit Schlanstedter wie mit Tick war mit dem Kompromiß-Wachstum der Rastatter verbunden, wobei das Wachstum während der kritischen Wochen zwischen Trieben und Früchten gleichmäßiger verteilt und offensichtlich bei den einzelnen Organen weniger kräftig ist.Zusammen mit dem gegensätzlichen Verhalten von A. pisum und früheren Arbeiten über die Phloemsaftzusammensetzung in Beziehung zu Pflanzenwuchs und Aphidenernährung weisen diese Beobachtungen darauf hin, daß der Phloemsaft für die Blattläuse bei der Rastatter am wenigsten nahrhaft sein dürfte und daß die Resistenz der Rastatter gegen Aphis fabae auf der Reaktion der geflügelten Migranten auf einige oberflächliche Pflanzeneigenschaften beruhen könnte, die mit Wachstumseigentümlichkeiten und der Saftzusammensetzung verbunden sind. Mögliche praktische Folgerungen werden kurz erwähnt.

     

Length heterogeneity in a region of the human ribosomal gene spacer is not accompanied by extensive population polymorphism

We carried out a restriction enzyme analysis of human ribosomal DNA structure on total placental DNA using the Southern (1975) method. Studies on a single individual using HindIII, PstI, HpaI and BglII revealed that a region of the non-transcribed spacer near the 3′ end of the 28 S gene was heterogeneous in size. Four fragment classes were detected in this individual. Adjacent classes differed in size from one another by about 0·8 × 10³ bases. Analysis of 19 additional placental samples and four cell lines revealed no fragment classes other than those detected in the original sample. Mixing experiments carried out with all 20 placental DNA samples provided further evidence for the discrete nature of the population polymorphism in the length of this region of the spacer. This finding contrasts sharply with the almost continuous nature of the population polymorphism in the length of a region of the non-transcribed spacer in Xenopus laevis.     

Intra-isolate heterogeneity of the ITS region of rDNA in Pythium helicoides

Heterogeneity of the rDNA ITS region in Pythium helicoides and the phylogenetic relationship between P. helicoides and closely related species were investigated. In PCR-RFLP analysis of the rDNA ITS region of six P. helicoides isolates investigated, including the type culture, intraspecific variation was found at the HhaI site. The total length of fragments was longer than before cutting, indicating sequence heterogeneity within isolates. Digestion of the cloned rDNA ITS region derived from seven isolates with HhaI revealed polymorphisms among and within single zoospore isolates, and variability of the region was also present among the clones derived from the same isolate. To test whether the rDNA ITS region of closely related species and other regions in the genome of P. helicoides are also variable, the rDNA ITS region of P. ultimum and the cytochrome oxydase II (cox II) gene encoded in mitochondria were sequenced. P. ultimum had little variation in the rDNA ITS region. The cox II gene sequences of both species revealed only a low intraspecific variability and no intra-isolate variation. In the phylogenic tree based on the rDNA ITS sequences, all clones of P. helicoides formed one large clade that was distinct from the clades comprising morphologically similar species, such as P. oedochilum and P. ostracodes, and was closely related to P. chamaehyphon rather than the other species.     

Nucleotide sequence of the split tRNAUAALeu gene from Vicia faba chloroplasts: evidence for structural homologies of the chloroplast tRNALeu intron with the intron from the autosplicable Tetrahymena ribosomal RNA precursor

Summary The gene encoding the tRNA _UAA ^Leu from broad bean chloroplasts has been located on a 5.1 kbp long BamHI fragment by analysis of the DNA sequence of an XbaI subfragment. This gene is 536 bp long and is split in the anticodon region. The 451 bp long intron shows high sequence homology over about 100 bp from each end with the corresponding regions of the maize chloroplast tRNA _UAA ^Leu intron. These conserved sequences are probably involved in the splicing reaction, for they can be folded into a secondary structure which is very similar to the postulated structure of the intron from the autosplicable ribosomal RNA precursor of Tetrahymena. Very little sequence conservation is found in the 5-and 3-flanking regions of the broad bean and maize chloroplast tRNA _UAA ^Leu genes.