【作者】 邓川；

【导师】 林辰涛；

【作者基本信息】 吉林大学 ， 植物学， 2013， 博士

【摘要】 甜高粱具有生物学产量高，抗逆性强等生物学特性，使之成为最具潜力的生物质能源作物之一。筛选和培育适宜边际土壤种植的抗旱耐盐碱甜高粱品种对于发展燃料乙醇产业具有重要意义。本研究选取29份不同基因型甜高粱和2份粒用高粱为试验材料，在吉林省西部、中部三个不同生境下［干旱胁迫（E1）、干旱与盐碱胁迫共存（E2）、正常（E3）］，根据甜高粱生物鲜重、生物干重、茎汁糖锤度及理论酒精产量等能源性状在不同生境下的表现，筛选抗旱耐盐碱基因型甜高粱。通过AMMI模型及双标图，分析了不同基因型甜高粱的能源性状在不同生境下的稳定性及适应性。研究了不同耐性基因型甜高粱在不同时期的生理生化指标变化，以期从多方面揭示甜高粱抗旱、耐盐碱机制，丰富甜高粱抗逆生理内容，为抗旱、耐盐碱品种选择和旱地、盐碱地甜高粱栽培提供理论依据。同时，以甜高粱幼穗作为外植体，对不同基因型甜高粱离体培养再生能力进行评价，研究了基因型和激素配比对愈伤组织的诱导和分化再生的影响，为进一步建立甜高梁高效遗传转化体系奠定基础。取得的主要研究结果如下：1.甜高粱抗旱耐盐碱种质资源筛选干旱和苏打盐碱胁迫共存下，甜高粱出苗受到抑制，平均出苗率为75.1%。甜高粱出苗率基因型间存在差异，表明不同基因型甜高粱的苗期耐盐碱性有一定差异。其中九甜4号具有较强的耐受性，耐性最差的为MN-3020、MN-2745。对三个生境下甜高粱的生物鲜重、生物干重、茎汁糖锤度和理论酒精产量等能源性状进行方差分析和多重比较分析，结果表明，基因型、环境和基因型与环境互作间均有显著差异。不同基因型甜高粱生物鲜重在E1生境下变异幅度为23.3t·hm^-2–101.9t·hm^-2，变异系数为17.43%；E2生境下为10.4t·hm^-2-54.8t·hm^-2，变异系数为49.90%；E3生境下为20.8t·hm^-2-101.2t·hm^-2，变异系数为13.93%。生物干重在E1生境下变异幅度为5.7t·hm^-2-23.8t·hm^-2，变异系数为9.12%；E2生境下为3.0t·hm^-2-13.4t·hm^-2，变异系数为11.71%；E3生境下为3.5t·hm^-2-23.8t·hm^-2，变异系数为11.16%。茎汁糖锤度在E1生境下变异幅度为12.9%-22.8%，变异系数11.84%；E2生境下为6.4%-19.1%，变异系数为17.50%；E3生境下为8.2%-18.4%，变异系数为9.18%；理论酒精产量在E1生境下变异幅度为1771L·hm^-2-9608L·hm^-2，变异系数为11.09%。E2生境下为577L·hm^-2-5454L·hm^-2，变异系数为13.36%。E3生境下为911L·hm^-2-9154L·hm^-2，变异系数为9.02%。结果表明，不同基因型甜高粱生物鲜重、生物干重、茎汁糖锤度及理论酒精产量在同一生境下表现均不同。在三个不同生境下，生物鲜重在E3表现最好，平均生物鲜重达55.8t·hm^-2。在E1表现次之。生物鲜重在E2生境下表现最差，仅为28.1t·hm^-2。生物干重在E3、E1表现最好，平均生物干重分别达13.6t·hm^-2和13.2t·hm^-2。生物干重在E2生境下表现最差，仅为7.2t·hm^-2。茎汁糖锤度在E1表现最好，平均茎汁糖锤度达18.2%。在E3和E2表现次之，分别为15.7%和15.2%。理论酒精产量在E3和E1最高，平均理论酒精产量达4864L·hm^-2和4854L·hm^-2。在E2生境下最低，仅为2426L·hm^-2。说明不同生境对生物鲜重、生物干重、茎干糖锤度和理论酒精产量等能源性状均有影响。通过AMMI模型及双标图，分析了不同基因型甜高粱的能源性状在不同生境下的稳定性及适应性。根据高产、高糖、稳定性高的筛选标准，结合不同基因型甜高粱的出苗率，与其它供试基因型相比，九甜杂3、散穗甜生物产量最高，茎汁糖锤度较高，在半干旱、盐碱地的酒精理论值分别为5454L·hm^-2和4605L·hm^-2；半干旱、非盐碱地酒精理论值可达到9608L·hm^-2和6961L·hm^-2。九甜杂3、散穗甜符合生物能源优良品种的要求，可作为生物能源型品种在吉林省西部半干旱的边际土壤上推广种植。九甜杂4出苗率高，生物产量和糖锤度均较高，但受环境影响较大，适宜在非盐碱地土壤上种植。九甜杂4在半干旱地区酒精理论值最高达到7761L·hm^-2。2.甜高粱抗旱耐盐碱生理生化响应研究选用4个抗旱、耐盐碱能力不同基因型的甜高粱（九甜杂3、散穗甜、MN-3018和MN-3382）为材料，测定甜高粱的在3个不同生境下苗期、拔节期、开花期和成熟期的生理指标。SOD、POD、CAT活性、叶片细胞膜透性相对电解质渗出率、脯氨酸和MDA含量在E3生境下最低，其次为E1，在E2生境下最高。4个基因型间各生理指标在不同生境下存在明显差异，九甜杂3和散穗甜升高或降低的幅度均大于MN-3018和MN-3382。以上试验结果表明，干旱、盐碱胁迫对质膜严重破坏，叶片细胞膜透性相对电解质渗出率增加，大量有毒物质产生，MDA含量升高。甜高粱通过提高体内抗氧化酶SOD、POD、CAT酶活和脯氨酸含量来调节对干旱、苏打盐碱逆境的适应性。不同基因型间耐受差异明显，通过生理指标的升高或者降低可判断基因型的抗性强弱。测定4个基因型的甜高粱在不同生境下茎秆的可溶性总糖、蔗糖、果糖和葡萄糖含量。结果表明，不同基因型间各糖分变化差异显著，干旱、盐碱胁迫对甜高粱茎秆的可溶性总糖、蔗糖、果糖和葡萄糖均有影响。3.甜高粱离体培养再生能力评价在设置的4种培养基上29个基因型甜高粱被诱导出愈伤组织，16个基因型甜高粱可分化再生出植株,不同基因型间愈伤组织诱导率及分化率有明显差异。不同激素浓度配比对愈伤组织诱导的影响有较大差别，且基因型和不同浓度配比之间存在相互作用。Y3培养基（激素配比为2,4-D3mg/L+KT0.5mg/L）诱导效果最好，其平均诱导率达40.1%。褐化现象贯穿着整个外植体成愈过程，不同基因型甜高粱褐化程度不同。综合出愈率、分化能力、褐化程度等几方面的表现，本试验认为基因型MN-3020、1788、九甜1号、吉甜3号和685-525的幼穗对组织培养的反应相对较好，可作为基因转化的首选材料。更多还原

【Abstract】 Sweet sorghum（Sorghum bicolor L.） is characterized by drought and floodresistance, salt tolerance and high efficiency of biomass accumulation. It is consideredto be one of the most promising bio-energy crops. Choosing drought-resistant andsalt-enduring varieties is the most economic, safe and effective method in improvingthe products and qualities of sweet sorghum. The experimental material comprising oftwenty-nine sweet sorghum genotypes（cultivars and experimental lines） and two grainsorghum. Screening of sweet sorghum germplasm tolerant to drought and saline-alkalistress under different three inhabits in west Jilin of China. This study elucidated thechanges of physiological and biochemical indices under drought and saline-alkalistress in sweet sorghum systematically and revealed that the drought-resistant andsalt-enduring mechanisms of the sorghum, so it could provide the theoretical basis inselecting the drought-resistant and salt-enduring varieties and planting on drought andsaline-alkali soil. Thirty-one diverse genotypes of sweet sorghum were evaluated anddetermined their callus induction and plant regeneration capabilities using immatureinflorescences as explants. This study has identified the sweet sorghum genotypeswith the best potential for callus production and whole-plant regeneration in culture.This information is essential for successful genetic transformation of sweet sorghum.The results are as follows:1. Screening of sweet sorghum germplasm tolerant to drought and saline-alkali stress.The sweet sorghum emergence averaged75.1%under saline-alkali stress. Thesignificant difference in emergence of different sweet sorghum genotypes existed.JiuTian4sweet sorghum showed the greatest emergence, while MN-3020andMN-2745had the lowest.Variance analysis and multiple comparisons showed significant difference in fresh biomass yields, dry biomass yields, Brix degree of juice, theoretical ethanol yields ofthe31sweet sorghum genotypes grown in three inhabits.The variability of fresh biomass yields was from23.3t·hm^-2to101.9t·hm^-2in E1inhabit, from10.4t·hm^-2to54.8t·hm^-2in E2inhabit and from20.8t·hm^-2to101.2t·hm^-2in E3inhabit. The coefficient of variation was17.43%,49.90%,13.93%,respectively. The variability of dry biomass yields was from5.7t·hm^-2to23.8t·hm^-2in E1inhabit, from3.0t·hm^-2to13.4t·hm^-2in E2inhabit and from3.5t·hm^-2to23.8t·hm^-2in E3inhabit. The coefficient of variation was9.12%,11.71%,11.16%,respectively. The variability of Brix degree of juice was from12.9%to22.8%in E1inhabit, from6.4%to19.1%in E2inhabit and from8.2%to18.4%in E3inhabit. Thecoefficient of variation was11.84%,17.50%,9.18%, respectively. The variability oftheoretical ethanol yields was from1771L·hm^-2to9608L·hm^-2in E1inhabit, from577L·hm^-2to5454L·hm^-2in E2inhabit, from911L·hm^-2to9154L·hm^-2in E3inhabit. The coefficient of variation was11.09%,13.36%,9.02%, respectively. Theresults showed fresh biomass yields, dry biomass yields, Brix degree of juice,theoretical ethanol yields of each sweet sorghum genotypes were different in the sameinhabit.Fresh biomass yields showed the highest yields under E3(averaged55.8t·hm^-2)and showed the lowest yields under E2(averaged28.1t·hm^-2). Dry biomass yieldsshowed the highest yields under E3(averaged13.6t·hm^-2) and was similar underE1(averaged13.2t·hm^-2).The lowest yields (averaged7.2t·hm^-2) showed under E2.Brix degree of juice showed the highest under E1（averaged18.2%）. E3was similar toE1（averaged15.7%,15.2%, respectively）. Theoretical ethanol yields showed thehighest yields under E3(averaged4864L·hm^-2) and was similar to E1(averaged4854L·hm^-2). E2showed the lowest yields (averaged2426L·hm^-2). Fresh biomass yields,dry biomass yields, Brix degree of juice, theoretical ethanol yields of the31sweetsorghum genotypes were different in three inhabits.The energy traits of31sweet sorghum genotypes were analyzed with AMMImodel and biplot using data from three inhabits, and then the stability and adaptabilityof different genotypes were evaluated. According to the high-yield, high-sugar, high stability screening criteria, genotypes of JiuTianⅢand SanSuiTian were suitable forplanting in saline-alkali soil. Theoretical ethanol yields were5454L·hm^-2and4605L·hm^-2, respectively under saline-alkali stress. The theoretical ethanol yields ofJiuTian3was7761L·hm^-2under drought stress.2. Physiological and Biochemical characteristic responses of drought andsaline-alkali stress resistance in different sweet sorghum genotypes.Four different genotypes, JiuTianⅢ, SanSuiTian, MN-3018and MN-3382, ofsweet sorghum were used as materials to assayed the comparison of physiologicalcharacteristic in various growth stages. The rate of relative electrolyte, content ofMDA, proline and activity of SOD, POD, CAT showed the lowest under E3andshowed the highest under E2. And physiological characteristic showed significantdifferences among the four sweet sorghum genotypes, the change of JiuTianⅢandSanSuiTian were higher than MN-3018and MN-3382. The change of physiologicalcharacteristic showed that plasma membrane serious damaged under drought,saline-alkali stress. The rate of relative electrolyte, content of MDA, proline andactivity of SOD, POD, CAT in leaves were all increased in order to adapt to stress.The change of physiological characteristic may be the indexes of screening sweetsorghum genotypes for drought, saline-alkali tolerance.Saline-alkali stress affects the total soluble sugar, sucrose, fructose and glucose instalks of sweet sorghum. There were significant differences in different genotypes.3. Evaluation of regeneration potential of sweet sorghum genotypes.Twenty-nine of31genotypes of sweet sorghum were able to form calli and16genotypes were able to regenerate whole plants. Variance analysis showed significantdifferences in callus induction and plant regeneration frequency among differentgenotypes. Variance analysis showed significant difference in callus inductionfrequency among different media, indicating that2,4-D and KT concentrationsaffected callus induction frequency. The highest callus induction rate was obtained onY3medium [MS medium+3mg l-12,4-D+0.5mg l-1KT] with an average inductionrate of40.1%. The amount of phenolic compounds produced varied widely among thedifferent genotypes. The sweet sorghum genotypes MN-3020,1788,JiuTianⅠ,JiTian Ⅲ, and685-525showed the best callus induction and regeneration capabilities inthese conditions.更多还原