Cadmium (Cd2+) is one of the major widespread environmental pollutants, and can cause serious problems to all organisms. Lead (Pb2+) is another wide spread dangerous heavy metal. Tobacco is a popular growing eco...Cadmium (Cd2+) is one of the major widespread environmental pollutants, and can cause serious problems to all organisms. Lead (Pb2+) is another wide spread dangerous heavy metal. Tobacco is a popular growing economic crop in China. Most tobacco growing region soils contain excessive Cd2+ and Pb2+. To assess anatomic changes of tobacco roots under Cd2+, Pb2+, and Cd2++pb2+ chronic stress, a pot experiment was carried out in field. The tobacco seedlings with 6 leaves were transplanted to pots in which soil was placed. The amounts of Cd2+ added to soil were 0, 3, 6, 10, 30, 60, and 100 mg kg-1 dry soil. The amounts of Pb2+ added to soil were 0, 150, 300, 450, 600, 750, and 1 000 mg kg-1 dry soil. The amounts of Cd2++Pb2+ added to soil were 0+0, 3+150, 6+300, 10+450, 30+600, 60+750, and 100+1000 mg kg-1 dry soil. The contents of Cd2+ and Pb2+ in root systems were determined by inductively coupled plasma, and the anatomical structure was studied by method of paraffin sectioning. The results revealed that the amounts of exchangeable Cd2+ and Pb2+ and carbonate bound Cd2+ and Pbz+ in soil increased with the amounts of Cd2+ and Pb2+ added to soil, and the contents of both Cd2+ and Pb1+ in roots were significantly increased along with stress time and the amounts of Cd2+ and Pb2+ added to soil. The growing of tobacco in Cd2+ and Cd2++Pb2+ polluted soil for 50, 100, and 150 d resulted in some abnormal extemal morphological and anatomical changes in ripe region of lateral roots. All the abnormal roots had abnormal vascular cylinders, and the ratio of abnormal extemal morphological and anatomical changes of roots positively correlated with the Cd2+ contents in roots and stress time. While, there were no abnormal external morphological and anatomical changes of roots under Pb2+ stress. It was suggested that Cd2+ stress could cause abnormal anatomic changes of roots, but Pb2+ stress could not.展开更多
Nicotine is a secondary substance synthesized in tobacco roots. In flue-cured tobacco planting, tobacco decapitation is an effective practice to promote nicotine biosynthesis by regulation of the redistribution of tot...Nicotine is a secondary substance synthesized in tobacco roots. In flue-cured tobacco planting, tobacco decapitation is an effective practice to promote nicotine biosynthesis by regulation of the redistribution of total nitrogen amounts. However, proteins relevant to nicotine synthesis in tobacco roots has not been identified and characterized yet. It is important to explore the regulation of nicotine biosynthesis in tobacco roots. To identify the proteins relevant to nicotine synthesis, the protein patterns in roots of flue-cured tobacco (cv. K326) before and after decapitation were analyzed. In the present study, the protein patterns in roots of flue-cured tobacco were analyzed by two-dimensional electrophoresis (2-DE), and the differentially-expressed spots were identified by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Paired comparison of 2-DE maps revealed 26 spots of differentially-expressed proteins in roots before and after decapitation. Furthermore, nine differentially-expressed spots were identified. There were four proteins which were enzymes possibly involved in nicotine biosynthesis. In addition, the roles of the four enzymes in nicotine biosynthesis were discussed in a putative network. Our results would contribute to the understanding of the regulation pathway of nicotine biosynthesis and further to the molecular manipulation on the nicotine contents in flue-cured tobacco.展开更多
Nitrogen(N), the major forms of which are nitrate(NO3–) and ammonium(NH4+), plays an important role in plant growth and mediation of root development. However, the role of auxin in root growth in response to differen...Nitrogen(N), the major forms of which are nitrate(NO3–) and ammonium(NH4+), plays an important role in plant growth and mediation of root development. However, the role of auxin in root growth in response to different NH4+/NO3– ratios remains unclear. Two tobacco cultivars(Nicotiana tabacum L.) were adopted in this study, which displayed variant growth features under the situations with sole NO3– nutrition ratio(NH4+/NO3– ratio: 0/100), low NO3– nutrition ratio(NH4+/NO3– ratio: 97/3), and optimal NH4+/NO3– ratio(50/50). We investigated the effects of the different NH4+/NO3– ratios on the formation and elongation of lateral roots(LRs), auxin concentration, DR5::GUS expression, 3 H-labeled indole acetic acid([3 H]IAA) transport, and the expression of six PIN genes in tobacco roots. We also examined the effects of exogenous auxin and a transport inhibitor on LRs growth. The results are shown as follows, compared to optimal N nutrition conditions, the biomass and nitrogen(N) accumulation were largely reduced by sole and low NO3– nutrition treatment in NC89, but no difference was observed in Zhongyan 100. In most cases, sole and low NO3– nutrition impaired the elongation and formation of firstorder lateral roots(1° LRs), only in NC89, thus reducing the root growth. IAA concentration and DR5::GUS expression levels decreased in roots when NC89 was subjected to sole and low NO3– nutrition media, suggesting that different NH4+/NO3– ratios affect the transport of auxin from leaves to roots. Results were similar following exogenous NAA application to low NO3– nutrition treated seedlings. Based on direct [3 H]IAA transport measurement, the transport of polar auxin from shoots to roots decreased due to low NO3– nutrition. PIN4 expression levels were markedly decreased in roots of NC89by sole and low NO3– nutrition, while they were unaffected in Zhongyan 100 roots. Overall, our findings suggest that LRs formation in tobacco seedlings is regulated by NH4+/NO3– ratios via modifying polar transport of auxin.展开更多
[Objectives]The species and distribution of root-knot nematodes in tobacco growing regions of Yunnan were investigated,in order to provide reference for comprehensive prevention of root-knot nematodes disease and toba...[Objectives]The species and distribution of root-knot nematodes in tobacco growing regions of Yunnan were investigated,in order to provide reference for comprehensive prevention of root-knot nematodes disease and tobacco variety layout. [Methods]Fifty-four samples of root-knot nematode host on tobacco crops were collected from Kunming,Qujing and Yuxi cities in Yunnan from August to October in 2015. The species of root-knot nematodes were identified by observing perineal patterns and molecular biological testing( PCR of ribosomal DNA of ITS region and sequence characterized amplified region,SCAR). [Results] There were 24 samples belonging to Meloidogyne arenaria,accounting for 44. 44%; seven M. incognita samples,accounting for 12. 96%; one M. javanica sample,accounting for 1. 85%. Eleven samples were compound populations of M. javanica and M. arenaria,accounting for 20. 37%. Five were compound populations of M. incognita and M. arenaria,accounting for 9. 26%. Three were compound populations of M. incognita and M. javanica,accounting for 5. 56%. Three belonged to compound population of M. incognita,M. javanica and M. arenaria,accounting for 5. 56%. [Conclusions] The species of root-knot nematodes in tobacco growing areas of Yunnan mainly include M. incognita,M. javanica and M. arenaria. M. arenaria is the dominant population.展开更多
In order to explore the differences of amino acids in root exudates and plant tissues, we selected four flue-cured tobacco varieties [ Yunyan 87 ( Ⅰ), K326 (Ⅱ), NC102(Ⅲ), Hongda (Ⅳ)] as the test materials ...In order to explore the differences of amino acids in root exudates and plant tissues, we selected four flue-cured tobacco varieties [ Yunyan 87 ( Ⅰ), K326 (Ⅱ), NC102(Ⅲ), Hongda (Ⅳ)] as the test materials and studied the components and mass fractions of amino acids in rhizosphere soil, non rhizosphere soil, roots and leaves via high performance capillary zone electrophoresis. Ten amino acids were detected out from four varieties, including histidine, lysine, tryptophan, phenylalanine, valine, threonine, proline, serine, alanine and glycine. The mass fractions of amino acids in leaves and roots were higher than those in rhizosphere soil and non rhizosphere soil. In rhizosphere soil and non-rhizosphere soil, the mass fractions of lysine, phenylalanine, threonine, proline and glycine in susceptible varieties ( Ⅲ, Ⅳ) were higher than those in resistant varieties ( Ⅰ, Ⅱ). In rhizosphere soil, the total amino acids successively were Ⅳ 〉 Ⅲ 〉 Ⅰ 〉 Ⅱ, and in non rhi- zosphere soil were Ⅲ 〉 Ⅳ 〉 Ⅰ 〉 Ⅱ. The total amino acids in roots of variety Ⅳ was the highest, up to 6 359. 195 μg/g. In leaves, the total amino acids in varieties I and If were higher than those in varieties IfI and IV. The results indicated that amino acids could secrete to rhizosphere via root secretion, and the compositions and mass fractions of amino acids varied with flue-cured tobacco varieties, thus the secretion amount of susceptible varieties was higher than that of resistant varieties.展开更多
基金supported by the State Tobacco Mo-nopoly Administration of China (10200201005)
文摘Cadmium (Cd2+) is one of the major widespread environmental pollutants, and can cause serious problems to all organisms. Lead (Pb2+) is another wide spread dangerous heavy metal. Tobacco is a popular growing economic crop in China. Most tobacco growing region soils contain excessive Cd2+ and Pb2+. To assess anatomic changes of tobacco roots under Cd2+, Pb2+, and Cd2++pb2+ chronic stress, a pot experiment was carried out in field. The tobacco seedlings with 6 leaves were transplanted to pots in which soil was placed. The amounts of Cd2+ added to soil were 0, 3, 6, 10, 30, 60, and 100 mg kg-1 dry soil. The amounts of Pb2+ added to soil were 0, 150, 300, 450, 600, 750, and 1 000 mg kg-1 dry soil. The amounts of Cd2++Pb2+ added to soil were 0+0, 3+150, 6+300, 10+450, 30+600, 60+750, and 100+1000 mg kg-1 dry soil. The contents of Cd2+ and Pb2+ in root systems were determined by inductively coupled plasma, and the anatomical structure was studied by method of paraffin sectioning. The results revealed that the amounts of exchangeable Cd2+ and Pb2+ and carbonate bound Cd2+ and Pbz+ in soil increased with the amounts of Cd2+ and Pb2+ added to soil, and the contents of both Cd2+ and Pb1+ in roots were significantly increased along with stress time and the amounts of Cd2+ and Pb2+ added to soil. The growing of tobacco in Cd2+ and Cd2++Pb2+ polluted soil for 50, 100, and 150 d resulted in some abnormal extemal morphological and anatomical changes in ripe region of lateral roots. All the abnormal roots had abnormal vascular cylinders, and the ratio of abnormal extemal morphological and anatomical changes of roots positively correlated with the Cd2+ contents in roots and stress time. While, there were no abnormal external morphological and anatomical changes of roots under Pb2+ stress. It was suggested that Cd2+ stress could cause abnormal anatomic changes of roots, but Pb2+ stress could not.
基金Natural Science Foundation of Henan Province, China (0624050013)Inovation Foundation of Tobacco Profession Cultivation Key Laboratory,China (06 TCIF 006)
文摘Nicotine is a secondary substance synthesized in tobacco roots. In flue-cured tobacco planting, tobacco decapitation is an effective practice to promote nicotine biosynthesis by regulation of the redistribution of total nitrogen amounts. However, proteins relevant to nicotine synthesis in tobacco roots has not been identified and characterized yet. It is important to explore the regulation of nicotine biosynthesis in tobacco roots. To identify the proteins relevant to nicotine synthesis, the protein patterns in roots of flue-cured tobacco (cv. K326) before and after decapitation were analyzed. In the present study, the protein patterns in roots of flue-cured tobacco were analyzed by two-dimensional electrophoresis (2-DE), and the differentially-expressed spots were identified by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Paired comparison of 2-DE maps revealed 26 spots of differentially-expressed proteins in roots before and after decapitation. Furthermore, nine differentially-expressed spots were identified. There were four proteins which were enzymes possibly involved in nicotine biosynthesis. In addition, the roles of the four enzymes in nicotine biosynthesis were discussed in a putative network. Our results would contribute to the understanding of the regulation pathway of nicotine biosynthesis and further to the molecular manipulation on the nicotine contents in flue-cured tobacco.
基金funded by the Agricultural Science and Technology Innovation Program, Chinese Academy of Agricultural Sciences (ASTIP-TRIC03)the Science Foundation for Young Scholars of Tobacco Research Institute of Chinese Academy of Agricultural Sciences (2018B01)+2 种基金the National Nature Science Foundation of China (3601818)the Liangshan Branch of Sichuan Tobacco Corporation, China (LSYC201805)the Hongyunhonghe Tobacco (Group) Co., Ltd., China (HYHH2016YL02)
文摘Nitrogen(N), the major forms of which are nitrate(NO3–) and ammonium(NH4+), plays an important role in plant growth and mediation of root development. However, the role of auxin in root growth in response to different NH4+/NO3– ratios remains unclear. Two tobacco cultivars(Nicotiana tabacum L.) were adopted in this study, which displayed variant growth features under the situations with sole NO3– nutrition ratio(NH4+/NO3– ratio: 0/100), low NO3– nutrition ratio(NH4+/NO3– ratio: 97/3), and optimal NH4+/NO3– ratio(50/50). We investigated the effects of the different NH4+/NO3– ratios on the formation and elongation of lateral roots(LRs), auxin concentration, DR5::GUS expression, 3 H-labeled indole acetic acid([3 H]IAA) transport, and the expression of six PIN genes in tobacco roots. We also examined the effects of exogenous auxin and a transport inhibitor on LRs growth. The results are shown as follows, compared to optimal N nutrition conditions, the biomass and nitrogen(N) accumulation were largely reduced by sole and low NO3– nutrition treatment in NC89, but no difference was observed in Zhongyan 100. In most cases, sole and low NO3– nutrition impaired the elongation and formation of firstorder lateral roots(1° LRs), only in NC89, thus reducing the root growth. IAA concentration and DR5::GUS expression levels decreased in roots when NC89 was subjected to sole and low NO3– nutrition media, suggesting that different NH4+/NO3– ratios affect the transport of auxin from leaves to roots. Results were similar following exogenous NAA application to low NO3– nutrition treated seedlings. Based on direct [3 H]IAA transport measurement, the transport of polar auxin from shoots to roots decreased due to low NO3– nutrition. PIN4 expression levels were markedly decreased in roots of NC89by sole and low NO3– nutrition, while they were unaffected in Zhongyan 100 roots. Overall, our findings suggest that LRs formation in tobacco seedlings is regulated by NH4+/NO3– ratios via modifying polar transport of auxin.
基金Supported by Specialized Research Fund for Doctoral Program of Colleges and Universities of the Ministry of Education(20135302110003)Backup Talent Cultivation Project of Young and Middle-aged Academic Technology Leaders in Yunnan Province(2009CI054)
文摘[Objectives]The species and distribution of root-knot nematodes in tobacco growing regions of Yunnan were investigated,in order to provide reference for comprehensive prevention of root-knot nematodes disease and tobacco variety layout. [Methods]Fifty-four samples of root-knot nematode host on tobacco crops were collected from Kunming,Qujing and Yuxi cities in Yunnan from August to October in 2015. The species of root-knot nematodes were identified by observing perineal patterns and molecular biological testing( PCR of ribosomal DNA of ITS region and sequence characterized amplified region,SCAR). [Results] There were 24 samples belonging to Meloidogyne arenaria,accounting for 44. 44%; seven M. incognita samples,accounting for 12. 96%; one M. javanica sample,accounting for 1. 85%. Eleven samples were compound populations of M. javanica and M. arenaria,accounting for 20. 37%. Five were compound populations of M. incognita and M. arenaria,accounting for 9. 26%. Three were compound populations of M. incognita and M. javanica,accounting for 5. 56%. Three belonged to compound population of M. incognita,M. javanica and M. arenaria,accounting for 5. 56%. [Conclusions] The species of root-knot nematodes in tobacco growing areas of Yunnan mainly include M. incognita,M. javanica and M. arenaria. M. arenaria is the dominant population.
基金Supported by Project of Yunnan Hongyun Honghe Tobacco(Group)Co.,Ltd.(KX140906)
文摘In order to explore the differences of amino acids in root exudates and plant tissues, we selected four flue-cured tobacco varieties [ Yunyan 87 ( Ⅰ), K326 (Ⅱ), NC102(Ⅲ), Hongda (Ⅳ)] as the test materials and studied the components and mass fractions of amino acids in rhizosphere soil, non rhizosphere soil, roots and leaves via high performance capillary zone electrophoresis. Ten amino acids were detected out from four varieties, including histidine, lysine, tryptophan, phenylalanine, valine, threonine, proline, serine, alanine and glycine. The mass fractions of amino acids in leaves and roots were higher than those in rhizosphere soil and non rhizosphere soil. In rhizosphere soil and non-rhizosphere soil, the mass fractions of lysine, phenylalanine, threonine, proline and glycine in susceptible varieties ( Ⅲ, Ⅳ) were higher than those in resistant varieties ( Ⅰ, Ⅱ). In rhizosphere soil, the total amino acids successively were Ⅳ 〉 Ⅲ 〉 Ⅰ 〉 Ⅱ, and in non rhi- zosphere soil were Ⅲ 〉 Ⅳ 〉 Ⅰ 〉 Ⅱ. The total amino acids in roots of variety Ⅳ was the highest, up to 6 359. 195 μg/g. In leaves, the total amino acids in varieties I and If were higher than those in varieties IfI and IV. The results indicated that amino acids could secrete to rhizosphere via root secretion, and the compositions and mass fractions of amino acids varied with flue-cured tobacco varieties, thus the secretion amount of susceptible varieties was higher than that of resistant varieties.
文摘为探索长期连作状态下烟草根系分泌物内含有的化感物质组成成分,设置石英砂栽培和连作土壤栽培2种烟草培养处理,对2种烟草不同生长时期的根系分泌物进行提取,利用超高效液相色谱串联质谱(ultra-high performance liquid chromatography tandem mass spectrometry,UHPLC-MS/MS)对提取的根系分泌物进行分析,通过数据库比对、差异分析和时间聚类分析等方法筛选连作烟草根系分泌物中潜在的化感物质。结果表明,共筛选出11类潜在化感物质,分别为生物碱、萜类、胺类、有机酸、苯酸及其衍生物、脂肪酸、酚类、氨基酸和多肽、香豆素、肉桂酸及其衍生物和其他。生物碱、萜类、胺类和有机酸4类化合物相对含量占所有化合物的73.3%。L-烟碱、熊果酸、β-氨基丙腈、5-羟基吲哚乙酸、早熟素Ⅱ、单乙基己基邻苯二甲酸、己内酰胺、姜酚、L-乳酸、邻苯二甲酸二乙酯和2,3-二氢-3,5-二羟基-2-氧代-3-吲哚乙酸甲酯是连作烟草根系分泌物中相对含量较高的潜在化感物质。研究结果为烟草根系化感物质的化感作用研究及缓解烟草连作障碍方法研究提供理论基础。