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.     

硝态氮对小白菜铬污染毒性的调控作用

通过盆栽和水培试验,探讨了硝态氮对小白菜铬污染毒性的调控作用。结果表明:外源Cr6 对小白菜铬吸收和积累具有明显的促进效应,抑制了小白菜对铁养分的吸收并降低了小白菜的硝酸还原酶活性;硝态氮可有效缓减Cr6 对小白菜吸收铁和硝酸还原酶活性的抑制作用,促进小白菜碳氮代谢和Vc的生物合成,并刺激小白菜生长。在相同铬污染条件下,土壤硝态氮的增加促进了外源Cr6 向有机态转化,且硝态氮有协同强化小白菜吸收Cr6 的效应。表明硝态氮在促进小白菜生长的同时,也促进了小白菜对Cr6 的吸收,提高了小白菜的铬累积水平。     

Influence of Soil Water Matric Potential and Hydraulic Conductivity on the Germination of Rape (Brassica napus L.)

The influence of matric potential and hydraulic conductivityon the water absorption and germination of rape (Brassica napusL.) was investigated by comparative studies in three seed-soilwater systems. In all systems the rate of germination and totalgermination were substantially retarded with decreasing matricpotential. Hydraulic conductivity of the soil system was demonstratedto be a limiting factor in the germination process. The effectsof matric potential and hydraulic conductivity on water absorptionfollowed closely those demonstrated for germination. The influenceof matric potential was shown to be comparable to that of osmoticpotential provided consideration was given to the hydraulicproperties of the soil-seed system.     

硅对受土壤中镉污染的白菜生长和抗胁迫能力的影响

在土培盆栽条件下,硅可提高白菜的地上部鲜重和茎鲜重,但根鲜重下降,叶鲜重略有下降。硅可抑制白菜吸收镉,在0.3、0.6、1.2mg·kg~(-1)镉水平下,施硅可显著降低白菜地上部的镉含量,并在一定程度上提高叶中过氧化物酶(POD)活性,但降低超氧化物歧化酶(SOD)活性。0.5、1.0 g·kg~(-1)硅可提高白菜的叶绿素含量和过氧化氢酶(CAT)活性,降低叶细胞膜透性,从而提高其对镉胁迫的耐受力。较高的硅浓度对植物有一定的毒性,叶绿素含量和CAT活性都下降,细胞膜透性也增加。     

Acute Toxicity of water-borne dimethylnitrosamine (DMN) to Fundulus heteroclitus (L.)

Acute (120h) toxicity of the water-borne carcinogen, dimethylnitrosamine (DMN) to adult Fundulus heteroclitus (L.) at 20°C and 20.2%₀ salinity in natural seawater was determined. The LC-50 values were 8300 mg/1 at 24 h, 5500 mg/1 at 48 h, 4700 mg/1 at 72 h, 3300 mg/1 at 96 h and 2700 mg/1 at 120 h.
Gross lesions consisted of peritoneal and cephalic ecchymoses. Microscopic lesions included necrosis and sloughing of the gastrointestinal mucosa, haemorrhage and necrosis of the liver, and necrosis of taste buds, with severity and incidence being a function of DMN concentration. Acute pathological effects of DMN on Fundulus heteroclitus conformed closely to those reported in other species exposed to this compound by respiratory, oral or parenteral route. DMN concentrations used in this experiment greatly exceeded those expected to be found under natural conditions, and this report is intended both to improve our understanding of DMN-induced pathology in marine fishes and to provide further evidence of the consistency of acute lesions induced by DMN, regardless of test organism or mode of exposure.     

Alleviation of Cadmium Toxicity in Brassica juncea L. (Czern. & Coss.) by Calcium Application Involves Various Physiological and Biochemical Strategies

Calcium (Ca) plays important role in plant development and response to various environmental stresses. However, its involvement in mitigation of heavy metal stress in plants remains elusive. In this study, we examined the effect of Ca (50 mM) in controlling cadmium (Cd) uptake in mustard (Brassica juncea L.) plants exposed to toxic levels of Cd (200 mg L⁻¹ and 300 mg L⁻¹). The Cd treatment showed substantial decrease in plant height, root length, dry weight, pigments and protein content. Application of Ca improved the growth and biomass yield of the Cd-stressed mustard seedlings. More importantly, the oil content of mustard seeds of Cd-stressed plants was also enhanced with Ca treatment. Proline was significantly increased in mustard plants under Cd stress, and exogenously sprayed Ca was found to have a positive impact on proline content in Cd-stressed plants. Different concentrations of Cd increased lipid peroxidation but the application of Ca minimized it to appreciable level in Cd-treated plants. Excessive Cd treatment enhanced the activities of antioxidant enzymes superoxide dismutase, ascorbate peroxidase and glutathione reductase, which were further enhanced by the addition of Ca. Additionally, Cd stress caused reduced uptake of essential elements and increased Cd accumulation in roots and shoots. However, application of Ca enhanced the concentration of essential elements and decreased Cd accumulation in Cd-stressed plants. Our results indicated that application of Ca enables mustard plant to withstand the deleterious effect of Cd, resulting in improved growth and seed quality of mustard plants.     

Phytoextraction Potential of Poplar (Populus alba L. var. pyramidalis Bunge) from Calcareous Agricultural Soils Contaminated by Cadmium

To investigate the phytoextraction potential of Populus alba L. var. pyramidalis Bunge for cadmium (Cd) contaminated calcareous soils, a concentration gradient experiment and a field sampling experiment (involving poplars of different ages) were conducted. The translocation factors for all experiments and treatments were greater than 1. The bioconcentration factor decreased from 2.37 to 0.25 with increasing soil Cd concentration in the concentration gradient experiment and generally decreased with stand age under field conditions. The Cd concentrations in P. pyramidalis organs decreased in the order of leaves > stems > roots. The shoot biomass production in the concentration gradient experiment was not significantly reduced with soil Cd concentrations up to or slightly over 50 mg kg^–1. The results show that the phytoextraction efficiency of P. pyramidalis depends on both the soil Cd concentration and the tree age. Populus pyramidalis is most suitable for remediation of slightly Cd contaminated calcareous soils through the combined harvest of stems and leaves under actual field conditions.     

Toxicity of heavy metals to Asellus aquaticus (L.) (Crustacea,Isopoda)

Effects of heavy metals on the isopod Asellus aquaticus (L.) are studied by static toxicity tests. Results demonstrate that the species is sensitive to Cd⁺², Cr⁺⁶, Cu⁺², Fe⁺³, Hg⁺², Ni⁺² Pb⁺² and Zn⁺², but the toxicity of each metal is different. Differences are also found between adults and between adults and juveniles. The comparative analysis of all data on the toxicity has been performed on the concentrations of metal ions and not on metal compound concentrations.Criteria for establishing water quality in order to guarantee protection of the environment are discussed.     

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.     

模拟大气细颗粒物中镉沉降对小白菜的毒性效应研究

为探究大气细颗粒物中镉(Cadmium,Cd)沉降对小白菜(Brassica rapa var.chinensis(L.)Kitamura)的毒性效应,本研究模拟不同浓度梯度大气细颗粒物中镉沉降(0、150、300和500μg/m3),分别代表对照、轻度、重度和严重污染,检测幼苗受胁迫后的生长状况、叶绿素相对含量(SP...     

Effect of Soil Organic Matter Content and pH on Toxicity of Cadmium to Paronychiurus kimi (Lee) (Collembola)

The heterogeneous physico-chemical properties of soil make it difficult to normalize the toxicological effects of metals in the Collembolan species Paronychiurus kimi (Lee). Furthermore, the physico-chemical properties of soil themselves can function as limiting factors that affect population fitness. In this study, the effects of soil organic matter (OM) and pH on the biological performance of P. kimi were assessed, and the influence of these properties on cadmium toxicity to P. kimi was also assessed at the individual and population levels. The OM and pH were found to significantly influence offspring production. All toxicological values of cadmium to P. kimi showed consistent patterns of increase with increasing OM and pH values, implying a decreased toxicity. The 28-d LC₅₀ values varied from 34.9 to 115.9 mg/kg, and the 28-d EC₅₀ and 28-d r_i=0 also varied depending on the OM and pH values. These findings indicate that the OM and pH values themselves are important factors in determining not only the biological performance of P. kimi but also the toxicity of cadmium to P. kimi on the individual and population levels. Therefore, when ecological risk assessments are conducted for a certain soil, pre-examinations of the effects of abiotic factors and the selection of an appropriate endpoint must be accomplished before establishing levels of chemicals of concern (COC) for a certain species.     

Resistance of rape (Brassica napus) to attack by the cabbage aphid (Brevicoryne brassicae L.)

Resistance to Brevicoryne brassicae L. attacks in a New Zealand variety of forage rape resulted from a combination of host non–preference and antibiosis. Using clonal plant material obtained from cuttings, only half the numbers of immigrant alate cabbage aphids that settled to reproduce on the susceptible rape were to be found on the resistant rape. The reproduction rate of these alates was about 12 % slower on the resistant plants than on the susceptibles and the young took about 13% longer to mature. Antibiosis then shortened the reproductive life of the apterae by one-third, reduced their fecundity by nearly 50 % and caused 40% mortality in their progeny. The over-all effect of this was considerable and under conditions specified in the text could result in the population on the resistant plants being about one-eighth that on the susceptible plants in under 1 month. The resistant plants lost their resistance on flowering and became normally susceptible to cabbage aphid attack. With maturity, however, the B. brassicae-resistant plants became slightly resistant to Myzus persicae.     

Reduction of Cadmium Availability to Tobacco (Nicotiana tabacum) Plants using Soil Amendments in Low Cadmium-contaminated Agricultural Soils: A Pot Experiment

Cadmium (Cd) concentration in field-grown tobacco leaves usually ranges from < 0.5 to 5 mg Cd kg^–1 dry matter (DM). Reducing bioavailability of soil Cd by adding amendments to the soil could be suitable to mitigate Cd uptake by tobacco plants. However, little is known on the effect of inorganic amendments on agricultural soils with low Cd concentrations. Therefore, we performed a pot experiment with tobacco plants that were grown during 56 days in two neutral to alkaline agricultural soils with low total Cd concentrations (soil 1 = 0.4, soil 2 = 0.7 mg kg^–1). Both soils were amended or not with 1 or 5% of sepiolite, zeolite, hydroxyapatite and apatite II™. Major and trace elements were measured in mid-stalk position leaves. Soil metals were measured in a DTPA soil extraction to assess the effect of the amendments on metal bioavailability. Some amendments significantly reduced Cd concentration in tobacco leaves, but the effect differed between the two soils tested. In soil 1, the use of zeolite at the 1% dose was the most efficient, reducing the average Cd concentration from 0.6 to 0.4 mg kg^–1. In soil 2, the 5% hydroxyapatite treatment led to the maximal reduction in Cd concentration (50%), with an average final Cd concentration in leaves of 0.7 mg kg^–1 (control: 1.5 mg kg^–1). There was a dose effect for some amendments in soil 2 (containing more Cd), suggesting a reduced efficiency of the amendment at the lowest addition rate. DTPA extractable Cd and Zn measured at the end of the pot experiment were correlated to the metal concentrations in tobacco leaves suggesting that (1) the reduction in leaf Cd concentration was due to a reduction in metal availability to tobacco and (2) DTPA may be a suitable extractant to estimate Cd availability to tobacco plants in these two soils. In addition, a batch experiment was performed with the same soils to test a larger number of amendments, including the four tested in the pot experiment. Results were compared to those of the pot experiment to assess whether a batch experiment may predict the efficiency of an amendment on a given soil. It gave results compatible with those from the pot experiment except for the sepiolite and highlighted the broad range of potential amendments available for heavy metal remediation in crop plants.     

Trematophoma lignicola (Petrak): A Potential Microbial Herbicide for Control of Amaranthus retroflexus (L.)

Trematophoma lignicola (Petrak), sprayed on leaves at 106 conidia ml -1 in oil emulsion, kills Amaranthus retroflexus plants after a 16 h dew period. Application as a soil drench kills seedlings of this species.     

Effects of Spray Application of Lanthanum and Cerium on Yield and Quality of Chinese Cabbage (Brassica chinensis L) Based on Different Seasons

The use of single light rare earth elements in agriculture was studied using the application of single rare earth elements (La and Ce) to Chinese cabbage (Brassica chinensis L.) in field plot experiments where the soil conditions remained the same but the season (spring or autumn) in which application occurred was altered. The results showed that the Chinese cabbage’s nutrition quality between the two seasons had obvious differences. When planted in the autumn, the soluble sugar and vitamin C content was higher, the titratable acid and nitrate content was lower, and the ratio of sugar to acid was higher, while when planted in spring, the situation was opposite. La or Ce treatments in spring and autumn promoted the growth of the Chinese cabbage, the fresh and dry weight of the stems and leaves increased, the ratio of dry to fresh weight increased, and the observed effects were greater in the Ce treatments than the La treatments. Moreover, the soluble sugar content increased and the titratable acid content decreased, which meant that the ratio of sugar to acid increased. There was an increase in the vitamin C and nitrate content in the spring Chinese cabbage, while there was a decrease in the vitamin C and nitrate content in the autumn. The levels of the heavy metals Cu, Zn, Cd, Pb, and Ni decreased. The La treatment had greater effects on the spring Chinese cabbage, while the Ce affected the autumn Chinese cabbage more.     

Varietal susceptibility of certain broad bean seeds to infestation with Callosobruchus maculatus (F.) and Callosobruchus chinensis (L.) (Coleoptera: Bruchidae)

Seeds susceptibility of eight broad bean varieties to Callosobruchus maculatus (F.) and Callosobruchus chinensis (L.) infestation were studied for the first time in free- and no-choice methods in the laboratory of Plant Protection Department, Faculty of Agriculture, Zagazig University. The relation between certain seed physical characteristics and some biological and and infestation parameters of both insects in the two studied methods were evaluated. None of these varieties were resistant to both insects, showing various levels of susceptibility. Biological and infestation parameters were significantly different among varieties except the developmental period. In free- choice method, Giza 3 was the most susceptible variety to both insects, since produced the highest progeny of 246.67 and 75.67 adults and susceptibility index (SI) of 10.25 and 7.42, respectively, while the least susceptible variety was Giza 716. In no– choice method, Nubaria 5 and Sakha 1 were the most susceptible varieties to C. chinensis, while Nubaria 3 and Giza 3 to C. maculatus. Differences between physical characters of varieties were significant. Seed hardness were correlated negatively and seed coat thickness positively with laid eggs, progeny and (SI) of both insects in free-choice method. Also seed coat thickness correlated positively with weight loss and seed damage (%) of C. chinensis and negatively of C. maculatus. To reduce seed losses the cultivation of the least susceptible variety (Giza 716) is encouraged and considered for breeding purposes to avoid insecticide usage.     

Characterization and functional analysis of a novel PCP gene BcMF5 from Chinese cabbage (Brassica campestris L. ssp. chinensis Makino)

Brassica campestris Male Fertile 5 (BcMF5), a novel member of the pollen coat protein class A (PCP-A) gene family, was identified from Brassica campestris L. ssp. chinensis Makino (Chinese cabbage-pak-choi). Temporal and spatial expression analysis showed that BcMF5 is a late-expressed PCP gene related to the process of determining pollen fertility. Functional analysis by hairpin RNA (hpRNA)-mediated RNA interference also showed that the expression of BcMF5 is inhibited, which resulted in the low germination ability of the pollen and also in an abnormality of the pollen exemplified by a collapsed germination furrow. This demonstrates that the expression of BcMF5 is closely related to the tapetum. Further, the expression profile of the BcMF5 promoter in Arabidopsis was also analyzed. This analysis indicated that the BcMF5 promoter began expression in the early stage of anther development and drove high levels of glucuronidase (GUS) expression in anthers, pollen, and the pollen tube in the late stage of pollen development, but did not drive any expression in petals, sepals, or pistils. Together with the functional analysis, the hypothesis that BcMF5 may have a sporophytic or gametophytic expression pattern is presented.     

Cloning and Characterization of the Microspore Development-Related Gene BcMF2 in Chinese Cabbage Pak-Choi (Brassica campestris L. ssp. chinensis Makino)

For the sake of providing some important information relevant to the study of the molecular mechanism of genic male sterility in plants, gene differential expression in flower buds at different developmental stages, as well as in rosette leaves, florescence leaves, and scapes was analyzed using cDNA amplified fragment length polymorphism (cDNA-AFLP) in the genic male sterile A and fertile B line of Chinese cabbage pak-choi. Following amplification of 125 pairs of primer combinations, 11 differential fragments were obtained, of which eight were from the B line and the other three were from the A line. Of 11 differential fragments, four were verified by Northern hybridization that were expressed preferentially in fertile flower buds. Results of GenBank BLAST showed that one fragment was with unknown function,whereas the other fragments have strong nucleotide sequence similarities with the polygalacturonase (PG)gene, the pectinesterase (PE) gene, and the polygalacturonase inhibitory protein (PGIP4) gene. Only fulllength cDNA from the differential fragment BcMF-A18T16-1 was amplified by rapid amplification of cDNA ends (RACE) and Northern analysis showed that this fragment was expressed only in medium and largesized flower buds of the B line. The full-length cDNA, designated as BcMF2 (Brassica campestris Male Fertile 2), was 1 485 bp long and was composed of a 1 263-bp open reading frame, which had 83% nucleotide similarity to a PG gene from Arabidopsis encoding polygalacturonase. Analysis of the basic structure of the protein revealed that it had one polygalacturonase active site (RVTCGPGHGLSVGS) at 256th site of amino acids and was classified as being a member of family 28 of the glycosyl hydrolases. The role of the BcMF2 gene on microspore development is discussed in the present paper.     

Cloning and Characterization of the Microspore Development-Related Gene BcMF2 in Chinese Cabbage Pak-Choi （Brassica campestris L. ssp. chinensis Makino）

For the sake of providing some important information relevant to the study of the molecular mechanism of genic male sterility in plants, gene differential expression in flower buds at different developmental stages, as well as in rosette leaves, florescence leaves, and scapes was analyzed using cDNA amplified fragment length polymorphism (cDNA-AFLP) in the genic male sterile A and fertile B line of Chinese cabbage pak-choi. Following amplification of 125 pairs of primer combinations, 11 differential fragments were obtained, of which eight were from the B line and the other three were from the A line. Of 11 differential fragments, four were verified by Northern hybridization that were expressed preferentially in fertile flower buds. Results of GenBank BLAST showed that one fragment was with unknown function, whereas the other fragments have strong nucleotide sequence similarities with the polygalacturonase (PG) gene, the pectinesterase (PE) gene, and the polygalacturonase inhibitory protein (PGIP4) gene. Only fulllength cDNA from the differential fragment BcMF-A 18T 16-1 was amplified by rapid amplification of cDNA ends (RACE) and Northern analysis showed that this fragment was expressed only in medium and largesized flower buds of the B line. The full-length cDNA, designated as BcMF2 (Brassica campestris Male Fertile 2), was 1 485 bp long and was composed ofa 1 263-bp open reading frame, which had 83% nucleotide similarity to a PG gene from Arabidopsis encoding polygalacturonase. Analysis of the basic structure of the protein revealed that it had one polygalacturonase active site (RVTCGPGHGLSVGS) at 256th site of amino acids and was classified as being a member of family 28 of the glycosyl hydrolases. The role of the BcMF2gene on microspore development is discussed in the present paper.     

Functional analysis of a novel male fertility CYP86MF gene in Chinese cabbage (Brassica campestris L. ssp. chinensis makino)

In our earlier work, a cytochrome P450 CYP86MF gene was isolated from floral bud of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa L.) by mRNA differential display PCR (DD-PCR) and rapid amplification of cDNA ends (RACE). To unravel the biological function of CYP86MF gene, the antisense fragment from the CYP86MF gene was transferred into Chinese cabbage pak-choi (B. campestris ssp. chinensis var. communis Tsen et Lee). Out of 22 plants transformed with the antisense gene constructed from the CYP86MF, 20 reached to flowering stage. Morphological investigations showed that the transgenic plants developed the normal floral organ. However, they remained self-infertile, even when artificial self-pollination was performed in the bud stage. Pollen germination test indicated that the pollen from the transgenic line TB-2 could not germinate normally. Further physiological, biochemical and cytological analyses showed that only significant difference was detectable in contents of the endogenous hormones, and a layer of unknown material adhered to the surface of microspore. The present studies thus provided valuable clues for understanding the biological function of the CYP86C subfamily genes. Furthermore, our studies also demonstrate a novel method for obtaining artificial male sterility line of Chinese cabbage.