Effects of field application of phosphate fertilizers on the availability and uptake of lead, zinc and cadmium by cabbage (Brassica chinensis L.) in a mining tailing contaminated soil

A field trial was conducted to evaluate the reduction of bioavailability of heavy metals including lead (Pb), zinc (Zn) and cadmium (Cd) in a soil contaminated by mining tailings in Shaoxing, Zhejiang, China. Three commercial phosphate (P) fertilizers including phosphate rock (PR), calcium magnesium phosphate (CMP), and single superphosphate (SSP) were applied to the plot at three P application rates, 50, 300, and 500 g/m2 with 9 treatments and control (CK). Plants, water soluble and exchangeable (WE) extra...

铬胁迫对青菜(Brassica chinensis L.)植物螯合肽含量及抗氧化特性的影响

选用不同耐铬胁迫青菜品种春油1号和青伏令,采用不同浓度Cr3+处理,测定青菜抗氧化酶活性及植物螯合肽含量等生理指标。结果表明,随着Cr3+处理浓度升高,两种青菜叶片抗氧化酶活性、酸溶性SH、植物螯合肽(PCs)及谷胱甘肽(GSH)含量均呈增加趋势(P<0.05),高浓度Cr3(+300 mg.L-1)处理下,耐铬胁迫较弱的青伏令叶片SOD、POD、CAT活性分别比对照增加了24.2%、41.7%、21.4%,耐铬胁迫较强的春油1号分别比对照增加了37.2%、70.7%、43.0%,并且PCs含量是对照的3.17倍,明显高于耐铬胁迫较弱品种青伏令(1.71倍)。Cr3+处理下耐铬胁迫较强的青菜品种春油1号表现出相对较高的抗氧化酶活力及非酶物质含量,不同品种青菜对铬胁迫的生理差异较大。

铜、锌污染对水稻土中油菜(Brassica chinensis L.)生长的影响及累积效应研究

通过盆栽试验,系统研究了Cu、Zn重金属污染对油菜(Brassica chinensis L.)生长、SOD保护酶活性、脯氨酸、可溶性糖及Cu、Zn在油菜体内吸收富集的影响。结果表明:Cu、Zn污染对油菜生物量和生理生化指标(SOD保护酶活性、脯氨酸和可溶性糖)均有影响。随着Cu、Zn质量分数的升高,植株干物重和SOD保护酶活性表现为先增后降;当Cu质量分数达到1 000 mg?kg-1时,油菜干物重和SOD酶活性分别比对照下降36.79%、66.67%;当Zn质量分数达到3 000 mg?kg-1时,油菜干物重和SOD酶活性分别比对照下降55.21%、82.05%。Cu污染下,油菜叶片脯氨酸质量分数逐渐下降,总可溶性糖质量分数表现为先增后降;Zn污染下,脯氨酸和总可溶性糖质量分数均表现为先增后降。Cu、Zn污染使油菜幼苗叶片及根中的Cu、Zn质量分数明显增加,根部的积累尤其明显。当Cu处理为1 000 mg?kg-1时,油菜茎叶和根中质量分数分别为57.6、424.162 mg?kg-1;当Zn处理为1 000 mg?kg-1时,油菜茎叶和根中质量分数分别为240、608.54 mg?kg-1。与Zn相比,Cu主要积累在油菜的根部,向茎叶迁移累积的量很少,并且随着添加质量分数的增加,茎叶吸收Cu的量变化不大。总之,重金属Zn相对于Cu更容易积累于油菜的地上部分,从而更易进入食物链,它们在油菜体内富集都对其生理生化和营养产生了明显毒害作用。

沼液对无土栽培小白菜(Brassica chinensis L.)产量及品质的影响初探

无土栽培条件下,初步研究了沼液和化学肥料组合施用对小白菜(Brassica chinensis L.)产量及品质的影响。结果表明,当所施用的沼液营养液肥配方为每升沼液营养液肥中含沼液250 mL,配入3.0g尿素、2.7 g磷酸二氢钾、1.1 g氯化钙、2.5 g硫酸镁和2 mL微量元素储备液(处理3),小白菜产量最高,为10.6克/株,而且,与单纯施用化学肥料营养液(对照)相比,小白菜品质指标硝氮、Vc、可溶性糖及叶绿素含量并没有下降。生产中,合理利用沼液栽培小白菜,对提高其产量和品质有积极的作用。

Cow manure and cow manure-derived biochar application as a soil amendment for reducing cadmium availability and accumulation by Brassica chinensis L. in acidic red soil

Organic amendment is a promising,in situ phytostabilization approach to alleviate the phytotoxic effects of heavy metal contaminated soils.The aim of this study was to evaluate the feasibility of cow manure（CM）and its derived biochar（CMB）as a soil amendment on cadmium（Cd）availability and accumulation in low and high Cd-accumulating cultivars of Brassica chinensis L.grown in an acidic red soil.CM and CMB were applied to Cd-contaminated acidic red soil at the rates of 0,3.0and 6.0%（w/w）.Application of CMB was significantly more effective than that of CM,as it reduced the availability of Cd in soil by 34.3–69.9%and its bioaccumulation in the low Cd accumulator,Aijiaoheiye 333,by 51.2 and 67.4%,respectively.The addition of CMB significantly increased the extractability and accumulation of trace metals（Zn,Mn,Fe,and Cu）by plants and improved plant biomass production.CMB application,combined with utilizing low Cd accumulating cultivars represents a new,sustainable strategy to alleviate the toxic effects on Cd and improve food safety.

Comparisons in subcellular and biochemical behaviors of cadmium between Iow-Cd and high-Cd accumulation cultivars of pakchoi （Brassica chinensis L.）

Subcellular distributions and chemical forms of cadmium （Cd） in the leaves, stems and roots were investigated in low-Cd accumulation cultivars and high-Cd accumulation cultivars ofpakchoi （Brassica chinensis L.）. Root cell wall played a key role in limiting soil Cd from entering the protoplast, especially in the low-Cd cultivars. The high-Cd cultivars had significantly higher leaf and stem Cd concentrations than the low-Cd cultivars in cell wall fraction, chloroplast/trophoplast fraction, organelle fraction and soluble fraction. In low-Cd cultivars, which were more sensitive and thus had greater physiological needs of Cd detoxification than high-Cd cultivars, leaf vacuole sequestrated higher proportions of Cd. Cd in the form of pectate/protein complexes （extracted by 1 tool. L~ NaC1） played a decisive role in Cd translocation from root to shoot, which might be one of the mechanisms that led to the differences in shoot Cd accumulation between the two types of cultivars. Furthermore, the formation of Cd- phosphate complexes （extracted by 2% HAc） was also involved in Cd detoxification within the roots of pakchoi under high Cd stress, suggesting that the mechanisms of Cd detoxification might be different between low- and high-Cd cultivars.

The Effect of the Novel "light fertilizer" on the Environment Factor in High Tunnel and Growth of Brassica Chinensis

[Objective] The aim was to study the properties of novel ＂light fertilizer＂, photo-conversion film, in order to evaluate its effect on the environmental factors and morphogenetic process of crops in the high tunnel. [Method] Photo-conversion film and Iongevous anti-dropping film were compared in terms of light, air and soil temperature utilizing the Facilities of leafy agriculture high tunnel intelligent monitoring system. Verifying the results by farmland experiment. [Result] Photo- conversion film indeed improved the light quality of high tunnel compared with the Iongevous anti- dripping film. The air and soil temperature was raised several degrees. Results of farmland experiment show that the average value of brassica chinensis fresh weight increased 19.15% compared to the control. [Conclusion] Photo-conversion film promotes more crop growth than Iongevous anti-dropping film due to improvement of light quality, air and soil temperature.

Protective Effects of Glycinebetaine on Brassica chinensis Under Salt Stress

Brassica chinensis L. were foliarly applied with glycinebetaine (GB), as this species is unable to synthesis GB and sensitive to osmotic stress such as salt. The exogenous GB was easily absorbed and transported by the leaf of B. chinensis . Its application (0-20 mmol/L) enhanced the plant tolerance to salt stress. The treatment of 15 mmol/L GB significantly decreased the Na + accumulation in leaf and root under NaCl stress. This difference in accumulating Na + and K + is caused by higher selectivity of root absorption. Furthermore, GB increased H +_ATPase activity of root plasma membrane evidently. This result strongly suggested that in root the decreased Na + accumulation was caused by the GB accumulation that enhanced the extrusion of Na + from the cell in some way through plasma membrane transporter, e.g. Na +/H + antiport driven by H +_ATPase. The GB application was also found to stabilize the plasma membrane, to decrease the loss of chlorophyll, and to stimulate the osmosis induced proline response under salt stress.

Research Progress of Heat-tolerance of Brassica campestris ssp. Chinensis

Brassica campestris ssp. chinensis, also known as non-heading Chinese cabbage, is an important vegetable widely distributed in southern China. High temperature is the most common adversity factor in vegetable production, because Brassica campestris ssp. chinensis is a thermophilic vegetable, which can＇t well grow at high temperature. In summer and autumn, high temperature stress would prevent Brassica campestris ssp. chinensis from growing and result in poor quality of its seedlings, seriously influencing yield and quality of the vegetable in later period. In this paper, present situation and latest advances in heat resistance research of Brassica campestris ssp. chinensis in recent years were reviewed at home and abroad. The research tendency in Bassica campestris ssp. chinensis was also described, providing reference for breeding of heat-resistant Brassica campestris ssp. chinensis.

固相萃取-高效液相色谱法检测上海青(Brassica chinensis L.)中鱼藤酮的残留动态

[目的]建立固相萃取-高效液相色谱-紫外（SPE-HPLC-UV）检测上海青中鱼藤酮的分析方法,对上海青中鱼藤酮的含量进行动态分析。[方法]样品先经乙腈提取,随后在乙腈提取液中加入氯化钠,盐析后过Florisil固相萃取柱富集净化,再用正己烷-乙醚（体积比3∶7）洗脱,最后以乙腈-水（体积比6∶4）为流动相,在290 nm下用高效液相色谱检测分析上海青中鱼藤酮的含量。[结果]该方法稳定可靠,检测物在0.05－10 mg/L范围内线性关系良好。在0.5－5 mg/kg范围内,平均加标回收率为85.9%－86.3%,检出限为0.05 mg/kg。田间推荐剂量和2倍推荐剂量分别施药,鱼藤酮在上海青中的原始沉积量分别为6.26、18.34 mg/kg。[结论]该化合物在田间降解速率快,在上海青中的半衰期分别为11.2、11.1 h。

鸡毛菜(Brassica rapa L.Chinensis Group.)的颜色变化动力学及两种货架期预测方法

实验测定了贮藏在278、283、288、293K四个温度下的鸡毛菜的颜色参数L*、-a*、b*、△E、h*和感官评价,并对L*和△E进行了动力学分析。结果表明,在实验温度范围内,鸡毛菜储藏的温度越低,颜色参数变化越慢。动力学分析显示,零级动力学比一级动力学更适合表现鸡毛菜L*和△E的变化规律。研究还采用Arrhenius方程对颜色变化速率常数k和温度T进行非线性拟合,得到L*和△E活化能Ea分别为82.09kJ/mol和100.26kJ/mol。最后得到依赖于时间、温度和颜色指标的鸡毛菜货架期预测方程(R2>0.95),从中根据不同的L*和△E终点可得到对应的货架期。就本研究而言,假定L*增加12%为终点时,鸡毛菜在四个温度下的预测货架期分别为11.8、5.5、2.6、1.3d。同时,Arrhenius方程与依赖于感官终点的动态颜色终点拟合方程结合预测的货架期曲线与感官寿命曲线则能得到更好的契合(两者绝对差值小于0.6d),两者结合能得到较为全面的货架期预测参数。

Effect of Combined Use of Boron and Manganese Fertilizers on the Nutritional Quality and Physiological Indices of Brassica campestris L.ssp.chinensis L.var.utilis

[Objective] The purpose of this study is to determine the effects of com-bined use of boron and manganese fertilizers on the nutritional quality and physio-logical indices of Brassica campestris. [Method] In the nutrient solutions for growing B. campestris by hydroponics, boric acid and manganese sulfate were added at 0.5, 2.5, and 7.5 mg/L respectively. Another treatment without boron and manganese was prepared as the control. Quality and physiological indices of B. campestris in the 10 treatments were measured. [Result] Boron and manganese shared obvious in-teraction in improving the quality and physiological indices of B. campestris. To cul-tivate B. campestris with high quality and strong resistance, the optimum concentra-tions of boron and manganese in the nutrient solution should be 2.5 mg/L boric acid and 2.5-7.5 mg/L manganese sulfate. [Conclusion] The findings wil provide refer-ence for studying effects of trace elements on nutrient composition of vegetables.

Genetic Inheritance and Molecular Marker of Clubroot Resistance Genes in Brassica campestris ssp. chinensis

Objective] This study was conducted to investigate the genetic inheritance of clubroot resistance in Chinese non-heading cabbage （Brassica campestris ssp. chinensis）. [Method] The clubroot resistance gene was introduced from a Brassica campestris ssp. pekinensis cultivar to non-heading Chinese cabbage, and the inheri-tance and molecular markers of clubroot resistance gene in parental lines, F1, F2 and BC1 of non-heading Chinese cabbage were studied through pathogen inoculation at seedling stage and ISSR-PCR. [Result] Clubroot resistance in non-heading Chi-nese cabbage was control ed by a single dominant gene. ISSR molecular markers with Bulk segregant analysis （BSA） found that primer-873 was linked to resistance gene, named CR-873, and the genetic distance between the marker and the resis-tance gene was 9.72 cM. [Conclusion] The results provide references for the molecular marker assisted breeding of non-heading Chinese cabbage.

Role of nitrification inhibitor DMPP(3,4-dimethylpyrazole phosphate) in NO^-_3-N accumulation in greengrocery( Brassica campestris L. ssp. chinensis ) and vegetable soil

The influence of nitrification inhibitor(NI) 3,4 dimethylpyrazole phosphate(DMPP) on nitrate accumulation in greengrocery( Brassica campestris L. ssp. chinensis ) and vegetable soil at surface layer were investigated in field experiments in 2002 and 2003 Results showed that NI DMPP took no significant effect on yields of edible parts of greengrocery, but it could significantly decrease NO - 3 N concentration in greengrocery and in vegetable soil at surface layer. In addition, NI DMPP could reduce the NO - 3 N concentration during the prophase stage of storage.

Effect of Nitrogen and Sulfur Supply on Glucosinolates in Brassica campestris ssp.chinensis

Glucosinolates （GSs） are a group of plant secondary metabolites containing abundant nitrogen （N） and sulfur （S） mainly in Brassica and have the beneficial effects on human health including anti-carcinogenic, cholesterol-reducing and other pharmacological effects. The objective of this study was to investigate the effect of different concentrations of N （5, 10, and 20 mmol L-a, denoted by N5, N10 and N20） and S （0,5, 1, and 2 mmol L^-1, denoted by S0.5, S1 and S2） on the yield and GSs in pakchoi （Brassica campestris L. ssp. chinensis var. communis） in hydroponics. Results showed that N10 and N20 significantly enhanced the yield compared with N5, however, N20 had a negative effect relative to N10. Only with N10 and N20 low S supply （S0.5） reduced the yield. The concentrations of aliphatic GSs, aromatic GS and total GSs were enhanced by N5 and indolyl GSs were enhanced by N20. S2 enhanced the concentration of individual GS and total GSs. The concentrations of indolyl GSs were maximized in N20S2 treatment, whereas the highest concentrations of aliphatic GSs, aromatic GS and total GSs were found in N5S2 treatment. Effects of N and S on aliphatic GSs were higher than on indolyl GSs. The results suggest that the accumulation of aliphatic GSs and aromatic GS could be enhanced by low N and high S and restricted by high N while that of indolyl GSs could be enhanced by high N and high S.

甘蓝型油菜BnaSLY1基因进化分析及功能研究

赤霉素调控植物表皮细胞增长、茎叶伸长以及株型建成。拟南芥SLY1属于F-box蛋白,它通过靶向泛素化赤霉素信号转导路径的负向调控因子——DELLA蛋白来影响植物生长发育,然而油菜中BnaSLY1的基因功能尚未揭示。本研究对BnaSLY1进行了表达特征和进化树分析,利用CRISPR/Cas9技术创制了BnaSLY1不同拷贝数的突变体,结合RNA-Seq技术对BnaSLY1的生物学功能及其对油菜生长发育的影响进行了研究。结果表明,甘蓝型油菜Westar中有2个SLY1同源拷贝BnaA01.SLY1和BnaA06.SLY1,它们表达模式基本相同,为组成型表达基因,其蛋白定位在细胞核,且在不同的油菜品种及十字花科植物间序列保守。与对照相比,单突bnaa01sly1和bnaa06sly1开花时间推迟,株高显著降低,而双突bnasly1还表现出深绿色光叶表型,叶片厚度增加,开花期比单突进一步推迟,株高也进一步降低。RNA-Seq结果显示,双突与Westar之间的差异表达基因显著富集在生长素信号转导路径以及蜡质合成通路,多个开花时间相关基因表达也发生显著变化。本研究表明, BnaSLY1除影响植物株高和开花时间等生长发育进程,还影响表皮蜡质合成,为探索赤霉素信号转导路径在甘蓝型油菜生长发育中的重要作用奠定了理论基础。

Synergistic defensive mechanism of phytochelatins and antioxidative enzymes in Brassica chinensis L. against Cd stress

Brassica chinensis L. was chosen and exposed to different concentrations of Cd exposure to evaluate its Cd-accumulating capacity and its potential cellular defensive mechanisms. Cd accumulation in the shoots and roots of B. chinensis was up to 1348.3±461.8 and 3761.0±795.0 mg per killogram of dry weight, respectively, under 200 μmol/L of Cd exposure. Increasing Cd accumulation in the plant was accompanied by rapid accumulation of phytochelatins (PCs), and the sequestration of Cd by PCs provided a primary cellular mechanism for Cd detoxification and tolerance of B. chinensis. Furthermore, malondialdehyde formation, hydrogen peroxide content and antioxidative enzyme activities such as superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase were observed in the shoots of Cd-stressed B. chinensis. Increasing enzyme activities in response to concentrations of 5 to 50 μmol/L Cd showed an efficient defense against oxidative stress, suggesting that the antioxidative system was a secondary defensive mechanism. These resulted in reduced free Cd damage and enhanced Cd accumulation and tolerance. Glutathione plays a pivotal role in these two detoxification pathways. In general, these results suggested that PCs and the antioxidative system are synergistic in combatting Cd-induced oxidative stress and that they play important roles in Cd detoxification of B. chinensis, and also give a deep understanding of the natural defensive mechanisms in plants under heavy metal stress.

Differential Expression Analysis of Genie Male Sterility A/B Lines in Chinese Cabbage-Pak-Choi (Brassica campestris ssp. chinensis Makino)

To determine differential expression of genie male sterility A/B lines in Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis Makino var. communis Tsen et Lee), we used the RNA fingerprinting technique, cDNA-AFLP analysis, in different developmental stages and different tissues. While no obvious differential expressions were observed in rosette leaves, florescence leaves, and scapes, some differential expressions were found in alabstrums of A/B lines and among leaves, scapes and alabstrums. We analyzed the al-abstrums collected in different developmental stages with 10 primer combinations. We got a unique band between middle size alabstrums and large alabstrums in B line in one of the ten pair primers, and in another one pair, one band reflecting a higher gene-expression level in A line than that in B line was obtained. No unique bands were found with the other primer combinations. The bands reflecting different gene-expression level were confirmed by Northern hybridization. The results indicated that cDNA-AFLP was a suitable tool for studying differential expression of genie male sterility in plants. SDS-polyacrylamide gel electrophoresis patterns of soluble proteins further verified the difference in A/B lines.

Effect of the Antisense BcMF12 Driven by the BcA9 Promoter on Gene Silencing in Brassica campestris L.ssp.chinensis

The study analyzed the silencing of BcMF12 gene regulated by BcA9 promoter in the transgenic pakchoi and confirmed the effect of antisense BcMF12 gene on the pollen development. A conserved BcMF12 gene fragment was amplified from the cDNA of flower buds in pakchoi （Brassica campestris L. ssp. chinensis, syn. B. rapa L. ssp. chinensis） and was fused to the anther specific BcA9 promoter. The plant antisense expression vector was constructed and then introduced into pakchoi via Agrobacterium-mediated transformation. The transgenic plants were screened by antibiotics and molecular analysis. PCR and Southern blot revealed that the antisense BcMF12-GUS fusion gene regulated by BcA9 promoter was integrated into transgenic plants. Northern blot suggested that the expression of BcMF12 gene was down-regulated significantly. The pollen germination rate of transgenic plants with antisense BcMF12 gene decreased as compared with that of the control plants. The expression of the gene BcMF12 related to the pollen development was inhibited by the antisense BcMF12 driven by BcA9 promoter, which consequently affected the pollen development in pakchoi.

CNGC20 plays dual roles in regulating plant growth and immunity in Brassica napus

Inflorescence architecture is determined by inflorescence length,branch angles and the density of siliques,which affects planting density,lodging resistance and mechanical operation in rapeseed.However,the molecular mechanisms controlling inflorescence architecture are poorly understood,restricting the progress of breeding varieties with ideal plant architecture in oilseed rape.In this study,we have identified and characterized a rapeseed inflorescence development mutant,reduced inflorescence length(ril),which exhibits determinate and shortened inflorescences,reduced plant height,compact branches,and increased silique density.Through BSA-seq and map-based cloning,we find that RIL encodes a cyclic nucleotide-gated channel 20(BnaA01.CNGC20).A substitution of proline at the 304th position to leucine(P304L)was identified in the conserved transmembrane domain of BnaA01.CNGC20.This P304L substitution neither affects the subcellular localization and self-assembly of BnaA01.CNGC20,nor disrupts the interactions with BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1(BAK1),which interacts with CNGC20 and phosphorylates it to regulate Ca^(2+)channel stability.However,the P304L substitution increases channel activity and Ca^(2+)influx,which in turn induces immune responses such as cell death,H2O2 accumulation,upregulation of pathogenesis-related genes,and pattern-triggered immunity.The enhanced immunity improves the resistance to Leptosphaeria biglobosa and Sclerotinia sclerotiorum.Transcriptome analysis further revealed that CNGC20 plays dual roles in regulating plant growth and immunity via the brassinosteroid and auxin signaling pathways.The findings in this study provide deeper insights into the intricate relationship between cytosolic Ca^(2+)level and plant development and immunity,as well as the trade-off between immunity and the performance of yield-related traits in the heterozygous plants(+/ril),which may serve as a guide for balancing yield and disease resistance in oilseed rape breeding.