【作者】 隆小华；

【导师】 刘兆普；

【作者基本信息】 南京农业大学 ， 植物营养， 2007， 博士

【摘要】 开发利用可再生资源与能源，大力发展生物质能源及生物基化学品，减少经济发展对石油的依赖，具有十分重要的战略意义。菊芋作为生物质能源植物，其生态适宜性极强，可利用荒地及滩涂土地资源进行规模化种植。本论文主要就国内菊芋基因型资源进行筛选后，从田间试验和温室试验对适合沿海滩涂种植、高产优质的基因型菊芋进行系统筛选，揭示不同基因型菊芋在海水胁迫下生理生化特性差异，阐明典型基因型菊芋耐海水机理，探索典型基因型菊芋在海水胁迫下代谢变化及其特征，同时进行海涂海水灌溉菊芋需肥规律研究，阐述其机制，并对海涂种植菊芋的生态安全性进行评价，为菊芋的综合利用集成技术的产业化资源平台建设提供支撑。试验结果如下：1)随海水浓度的增加，各基因型菊芋的根和地上部干重的增长速率均降低，各基因型菊芋叶片MDA含量、膜透性及地上部和根Na⁺和Cl^-含量均增高，但南芋1号和8号表现为更耐海水。菊芋幼苗地上部和根鲜重和干重与根部Na⁺含量呈极显著负相关，地上部和根鲜重与地上部和根部Cl^-含量呈显著负相关，地上部和根干重与地上部和根部K⁺含量呈显著正相关。应用幼苗叶片过氧化物酶同功酶电泳技术进行基因型亲缘关系检测表明，8个基因型菊芋过氧化物酶同功酶表现出明显的谱带差异。根据菊芋在海水处理下的表现分为3种类型：高耐海水型，中耐海水型和低耐海水型。在评价不同基因型菊芋对海水的响应时，地上部和根鲜重、干重及含水率的下降幅度及Na⁺、K⁺和Cl^-含量可作为主要的生理指标。2)在海涂以不同基因型菊芋（Helianthus tuberosus L．）为材料进行田间试验结果表明，南芋2号根和地上部生物量较其他基因型大，南芋5号和南芋3号株高在各浓度海水灌溉下均显著高于其他菊芋基因型，而茎粗在各处理下变化不一致，在30％海水灌溉下，南芋1号根和地上部物质积累未受到抑制作用，其他基因型菊芋的根和地上部物质积累受到了一定抑制。各基因型菊芋块茎产量差异较显著，在30％海水灌溉下，南芋2号产量显著高于其他基因型菊芋，各基因型菊芋块茎单重在各处理下差异也较显著，南芋1号和2号块茎单重最大，各基因型菊芋块茎总糖和菊糖含量差异较显著。随海水浓度的增加，各基因型菊芋根、茎和叶的Cl^-和Na⁺含量均增加，但基因型间差异较显著，叶片的Na⁺含量显著低于根和茎的Na⁺含量。从生物积累量和块茎产量、总糖和菊糖含量及离子分布看，南芋1号和南芋2号较其他基因型更适合在海涂利用适当浓度海水进行灌溉种植。3)各基因型菊芋叶片SOD、POD、CAT活性及MDA含量在各浓度海水下处理下变化不一致，且随时间延长变化也不一致，从叶片MDA含量看，南芋1号在海水处理下膜脂过氧化程度较南芋7号和南芋6号小，受伤程度最低。随海水浓度的增加，各基因型菊芋叶片可溶性糖和脯氨酸含量变化不一致，且随时间延长变化也不一致。海水处理对各基因型菊芋幼苗生理代谢特征有影响，但对各基因型菊芋幼苗的影响不一致，南芋1号表现为更耐海水。4)采用X-射线微区分析技术研究了0％，15％和30％海水处理对较耐海水基因型南芋1号和低耐海水基因型南芋7号幼苗根系、茎杆、叶片横切面不同细胞离子分配的影响。结果表明，海水胁迫导致两基因型菊芋幼苗体内Na⁺和Cl^-含量显著上升，但两基因型间有差异，南芋1号体内Na⁺峰值较南芋7号高，占离子总量的百分比也比南芋7号高，在30％海水处理下，两基因型幼苗体内Na⁺和Cl^-含量占总离子百分比50％以上，南芋1号茎中柱薄壁细胞高达80％以上。各处理下K⁺、Ca²⁺和Mg²⁺峰值变化不明显，但在15％和30％海水处理下，其占离子总量百分比较0％海水处理均降低。5)．透过透射电镜观察海水处理对较耐海水基因型南芋1号和低耐海水基因型南芋7号幼苗叶绿体超微结构的影响。海水胁迫导致两基因型菊芋幼苗叶绿体片层结构模糊、解体，边缘膜模糊，被膜破损，内部结构破坏，而南芋1号较南芋7号受损程度低。6)利用高效液色谱．电化学相（库仑电极）阵列检测技术检测海水处理下不同处理时间后菊芋体内绿原酸的变化情况及其他小分子物质的差异显示。随处理时间的延长，0％海水处理下氯原酸含量变化不显著，而15％和30％海水处理下氯原酸含量变化显著15％海水处理下，在1h时较2h和3h时高，而30％海水处理下在3h时较1h、2h和6h时高。海水处理后菊芋幼苗体内产生的其他一些未知的小分子物质尚有待定性和进一步考察其变化规律。7)在山东莱州海涂采用正交试验设计进行田间试验结果表明，各浓度海水浇灌下，随着施氮、磷量的增加菊芋主茎普遍增长和增粗。经过对海水与N肥及P肥的交互作用分析，可以看出W₂N₃和W₂P₃是优化组合；处理因子分析表明，影响菊芋产量的主要因素是不同浓度海水灌溉，N肥和P肥次之，其优化组合为W₂N₃P₃。8)增加磷、氮素浓度后，能显著缓解海水胁迫的抑制作用，显著增加菊芋幼苗叶片脯氨酸和可溶性糖含量，增强菊芋幼苗叶片SOD、POD和CAT活性；降低了MDA含量和膜透性；降低地上部和根Na⁺和Cl^-含量，增加K⁺、Ca²⁺和Mg²⁺含量。说明磷、氮素能够改善菊芋幼苗的营养状况，同时能够增强其抗盐性，且随海水浓度的增加其效应越明显。9)大田小区海水灌溉实验中，0-5cm土层盐分变化剧烈，在菊芋整个生育期降雨量为515 mm的情况下，至收获期时，各处理均能降至灌海水前的水平。75％海水灌溉，5-40 cm层土壤盐分呈积累趋势，次生盐溃化明显；50％海水灌溉后轻微积盐，若无充沛的雨水淋洗则须结合一定的农业措施以防造成次生盐害；25％海水浓度在灌溉定额为1600 m³hm^-2时能保持土壤盐分的盈亏平衡。海水灌溉后，Na⁺和Cl^-主要分布在5-20 cm土层，而Ca²⁺和Mg²⁺主要在20-40 cm土层。更多还原

【Abstract】 It is significative to exploit regeneration resource and energy, such as biologic-energy and biologic-chemistry matter, to decrease the requirement of petroleum for economy development. Helianthus tuberosus adapts many kinds of environment, especially can be planted in badlands and mudflats along the coast.Genotype varieties of H. tuberosus were used to select systemicly high yield and quality one to plant in mudflats along the coast in field and greenhouse experiments in the study. The characteristics on physiology and biological-chemistry of H. tuberosus under seawater stress were investigated. The mechanisms of tolerance-seawater and metabolize of representative H. tuberosus were studied. The disciplinarian of fertilizer needed for H. tuberosus and zoology security for mudflats irrigated with seawater were also researched. The results were below:1) Compared with the control, the growth speed of fresh and dry weight of shoots and roots of eight H. tuberosus genotype seedlings treated with 15% and 30% seawater all decreased. The MDA content and electrolyte leakage in leaves and contents of Na⁺ and Cl^- in the shoots and roots increased with seawater concentration increasing. The Nanyu No.1 and Nanyu No.8 were more seawater tolerant than other genotype. There was significantly negative correlation between the fresh and dry weight of shoots and roots and the contents of Na⁺ in the roots. And there was significantly negative correlation between the fresh weight of shoots and roots and the contents of Cl^- in shoots and roots. There was significantly positive correlation between the dry weight of shoots and roots and the contents of K⁺ in shoots and roots. Based on those, the H. tuberosus of eight genotypes tested could be divided into three groups, such as high seawater-tolerance, intermedia seawater-tolerance and low seawater-tolerance. The the katabatic scopes of fresh weight and dry weight and contents of Na⁺, K⁺ and Cl^- in the shoots and roots could be used as physiological indices for selecting different tolerant-seawater genotype H. tuberosus. There was intensity difference of isoperoxidase bands by the in polyacrylamide gel electrophoresis to check the descendiblity relation of genotypes.2) Field experiments were carried out to study effects of seawater irrigation on yield compositions and ion distribution of different genotypes of H. tuberosus growing in coastal mudflat along semiarid regions in 2005. Nanyu No.2 was higher than the others in biomass of the roots and aerial parts, and. Nanyu No.5 and Nanyu No.3 were taller than the others in all treatments, but the effect on diameter of the main stems of the plants varied sharply with treatment. In the treatment of irrigation with 30% seawater, Nanyu No.1 was not affected in biomass accumulation in roots and aerial parts, but the others to a varying extent. The plants differed sharply in yield between genotypes. Nanyu No.2 was the highest when irrigated with 30% seawater, and so did they in single tuber weight, total sugar and inulin content. Nanyu No.1 and Nanyu No.2 were the highest in unit tuber weight. Concentrations of Cl^- and Na⁺ in roots, stems and leaves of all genotypes increased with the seawater concentration increasing, but significant differences did exist between genotypes. Na⁺ content was lower in leaves than in roots and stems. Judging by biomass accumulation, yield of tubers, total sugar and inulin contents in tuber, and ion distribution, Nanyu No.1 and Nanyu No.2 were more adaptive to planting in coastal mudflat along semiarid regions with seawater irrigation than other genotypes.3) The changes of activities of SOD, POD and CAT and contents of MDA of three H. Tuberosus genotypes seedlings leaves of seawater-tolerance Nanyu No.l, intermedia seawater-tolerance Nanyu No.6 and low seawater-tolerance Nanyu No.7 were different under seawater treatments. Nanyu No.1 was injured less under seawater treatments than Nanyu No.7 and Nanyu No.6 judged by lipid peroxidation degree in leaves. The contents of proline and soluble sugar of H. Tuberosus genotypes seedlings leaves were different under seawater treatments. And the contents of proline and soluble sugar both changed with time lasting. There were different effects on the physiological and metabolize characteristics of H. Tuberosus genotypes seedlings under seawater treatments. Nanyu No.l was more seawater-tolerant than Nanyu No.7 and Nanyu No.6.4) The distributions of ions in the cells of roots, stems and leaves of seawater-tolerance Nanyu No.1 and low seawater-tolerance Nanyu No.7 were analyzed by X-ray microanalysis technique. The contents of Na⁺ and Cl^- in the cells of roots, stems and leaves increased with seawater concentration increasing. But there was difference between the Nanyu No.1 and Nanyu No.7. The Na⁺ peak value of Nanyu No.1 was higher than Nanyu No.7. And the Na⁺ percent in the ions of Nanyu No.1 was also higher than Nanyu No.7. The Na⁺ and Cl^- percents in the ions of the two genotypes were over 50%, even over 80% in the stelar parenchyma cell under 30% seawater treatment. The peak values of K⁺, Ca²⁺ and Mg²⁺ did not change significantly under different treatments. But K⁺, Ca²⁺ and Mg²⁺ percents in the ions under 15% and 30% seawater treatments all were lower than under 0% seawater treatment.5) Chloroplast ultrastructures of Nanyu No.l and Nanyu No.7 under 0%, 15% and 30% seawater treatments were observed by clairvoyant micrograph. The configuration of chloroplast was blurry and disaggregation under seawater treatment. And the membrane was dilapidated. The configuration was also wrecked. But the breakage of Nanyu No.l chloroplast was lower than Nanyu No.7.6)The concentrations of chlorogenicacid and low molecular weight compounds were determined by high performance liquid chromatography with a coulometric array detector （HPLC-CAD） . The concentrations of chlorogenicacid did not change significantly under 0% seawater treatment along the time lasting. While it changed significantly under 15% and 30% seawater treatment along the time lasting. It was higher after treatment for 1 hour than for 2 and 3 hours under 15% seawater treatment. While it was higher after treatment for 3 hour than for 1,2 and 6 hours under 30% seawater treatment. Some other compounds was observed as well during the treatment. They are believed to below molecular weight compounds and subject to further investigations.7)The field experiment was also carried out in seashore in Laizhou, Shandong Province to study the effect of salt and fertilizer application coupling under irrigation of different concentration seawater on H. tuberosus by orthogonal design. It was that W₂N₃ and W₂P₃ were the optimized combinations through the analysis of alternant effects of seawater, nitrogen and phosphorus. The main element that affected the yields was seawater, then fertilizers of N and P were subordinate. And the best combination was W₂N₃P₃.8) The fresh weights of roots and aerial parts increased with the concentration of phosphorus supplementation. The trends of dry weights of roots and aerial parts resembled the trends of fresh weights with the same treatments. The contents of proline and soluble-sugar increased with phosphorus supplementation. The activities of SOD, POD and CAT all significantly stimulated with phosphorus supplementation. The MDA content and ELP decreased with phosphorus supplementation. And the contents of Na⁺ and Cl^- decreased with phosphorus supplementation. Compared with the control, the contents of K⁺, Ca²⁺and Mg²⁺of the aerial part and root increased with phosphorus supplementation. Phosphorus enrichment of the seawater ameliorated the toxicity of seawater in H. tuberosus seedlings. Nitrogen supplementation of the seawater resulted in increasing fresh and dry weight of shoot and roots when compared with seawater treatment without N supplementation. Nitrogen supplementation of the seawater significantly enhanced the activities of antioxidant enzymes in leaves compared to the seawater treatments alone. Addition of N to seawater enhanced the contents of proline and soluble-sugars in the leaves, K⁺ and total-N of aerial parts and roots. N supplementation resulted in a declined concentrations of Na⁺ and Cl^- in aerial parts and roots of seawater-stressed plants. Nitrogen enrichment of the seawater ameliorated the toxicity of seawater in H. tuberosus by improving the antioxidative enzymes, accumulating of proline and soluble-sugars and altering the distribution of inorganic ions.9) Distribution and migration of Na⁺, Cl^-, Ca²⁺, Mg²⁺, K⁺ in 0-40cm soil irrigated by seawater were investigated. There was 515 mm rainfall in the whole postemergence. Though content of salinity in 0-5cm layer changed drastically, it reached the controlled level （no irrigation） when if. tuberosus were harvested. Salinity was accumulated conspicuous in 0-40cm soil layer and would brought on secondary salinization when the seawater irrigation concentration was 75%. Some agricultural measures must be afforded to stand off secondary salinization if no enough rainfall. There was a significant change of salinity in plough layer soil when irrigated by 25% seawater. Na⁺ and Cl^- were primarily distributed in 5-20cm layer, and Ca²⁺ and Mg²⁺ in 20-40cm with seawater irrigation.更多还原