甘蔗(Saccharum officinarum L.)品种遗传差异的AFLP分子标记分析

采用AFLP技术 ,从 6 4对引物中筛选出 5对带型分布均匀、多态性丰富且分辨能力强的引物 ,对两个在生产上广泛推广应用的甘蔗品种的AFLP指纹图谱进行分析 ,计算了两品种间的遗传相似性和遗传距离。结果表明 ,5对引物在 2个甘蔗品种间均存在显著的差异 ,其中多态性位点占总扩增位点的 10 2 % ,区分率达 10 0 %。这对应用AFLP技术鉴定甘蔗品种的真伪提供了依据。本文还对甘蔗AFLP分析体系的关键技术以及银染的安全性和可靠性进行了讨论。

Radiation-induced in vitro mutagenesis system for salt tolerance and other agronomic characters in sugarcane(Saccharum officinarum L.)

Gamma ray-induced in vitro mutagenesis and selection for salt(NaC l) tolerance were investigated in sugarcane(Saccharum officinarum L.). Embryogenic callus cultures were irradiated(10 to 80 Gy) and subjected to in vitro selection by exposure of irradiated callus to NaC l(0, 50, 100,150, 200, and 250 mmol L-1). Increasing NaC l concentrations resulted in growth reduction and increased membrane damage. Salt-selected callus lines were characterized by the accumulation of proline, glycine betaine, and Na+and K+concentration. Higher accumulation of proline and glycine betaine was observed in NaC l stressed callus irradiated at 20 Gy. Na+concentration increased and K+concentration decreased with increasing salt level. Irradiated callus showed50–60% regeneration under NaC l stress, and in vitro-regenerated plants were acclimatized in the greenhouse, with 80–85% survival. A total of 138 irradiated and salt-selected selections were grown to maturity and their agronomic performance was evaluated under normal and saline conditions. Of these, 18 mutant clones were characterized for different agro-morphological characters and some of the mutant clones exhibited improved sugar yield with increased Brix%,number of millable canes, and yield. The result suggest that radiation-induced mutagenesis offers an effective way to enhance genetic variation in sugarcane.

Expression of the Grifola frondosa Trehalose Synthase Gene and Improvement of Drought-Tolerance in Sugarcane （Saccharum officinarum L.）

海藻糖是在生物结构的稳定作为杀虫撒布剂工作并且在有机体提高应力公差到不能生活的压力的葡萄糖的非减少的 disaccharide。我们这里报导 Grifola frondosa 海藻糖 synthase (TSase ) 的表示为在甘蔗(Saccharum officinarum L.) 改进干旱忍耐的基因。transgene 的表示在 CaMVSSS 倡导者的二个双人脚踏车拷贝的控制下面并且由 Agrobacterium tumefaciens EHA105 转了进甘蔗。转基因的植物积累了海藻糖的高水平，直到 8.805-12.863 mg/g 新鲜重量，而它在无法发现的水平 innontransgenic 植物是在场的。转基因的植物与 Escherichia coliTPS (trehalose-6-phosphatesynthase ) 转变了或 TPP (trehalose-6-phosphate 磷酸酶) 严重地被阻碍并且让根，这被报导了词法改变。有趣地，我们的转基因的甘蔗植物没在这块地里有明显的词法变化和没有生长抑制。海藻糖累积 in35S-35S：TSase 植物导致了增加的干旱忍耐，由干旱和干旱显示出生理的索引例如界限浇 / 释放水，质膜渗透， malondialdehyde 内容， and b 满足的叶绿素，和草皮的活动和切除叶子的邮政部门的率。这些结果建议与 TSase 基因转变的转基因的植物能积累海藻糖的高水平并且提高了忍耐到干旱。

Genetic analysis for brix weight per stool and its component traits in sugarcane (Saccharum officinarum)

Brix weight per stool (BW) of sugarcane is a complex trait, which is the final product of a combination of many components. Diallel cross experiments were conducted during a period of two years for BW and its five component traits, in- cluding stalk diameter (SD), stalk length (SL), stalk number (SN), stalk weight (SW), and brix scale (BS) of sugarcane. Phenotypic data of all the six traits were analyzed by mixed linear model and their phenotype variances were portioned into additive (A), dominance (D), additive×environment interaction (AE) and dominance×environment interaction (DE) effects, and the correlations of A, D, AE and DE effects between BW and its components were estimated. Conditional analysis was employed to investigate the contribution of the components traits to the variances of A, D, AE and DE effects of BW. It was observed that the heritabilities of BW were significantly attributed to A, D and DE by 23.9%, 30.9% and 28.5%, respectively. The variance of A effect for BW was significantly affected by SL, SN and BS by 25.3%, 93.7% and 17.4%, respectively. The variances of D and DE effects for BW were also significantly influenced by all the five components by 5.1%～85.5%. These determinants might be helpful in sugarcane breeding and provide valuable information for multiple-trait improvement of BW.

Implementation and Evaluation of Certain Properties of a Polymer Matrix Composite Material Reinforced by Fibrous Residues of <i>Saccharum officinarum</i>in View of an Applicability Orientation

This study examines the implementation and characterization of a polymer matrix composite material reinforced by sugarcane residues. The aim of the study is to enhance the abundantly produced sugarcane bagasse in the form of residues in the processing plants of said sugar cane. The composite material developed takes into account the size parameters and mass load rate of the reinforcement, the variations of which are between 2.5 mm and 4 mm respectively for the first parameter and 10% to 25% for the second. The load on the test tube during the test has a random orientation. The composite is polyester matrix. The cold compression moulding technique was used in the production of the various samples. Physical properties such as the rate of water absorption and the density of the composite are assessed. The three-point bending tests are carried out with the aim of inducing Young’s modulus from the elaborate samples. Analysis of physical properties shows that water absorption increases with the rate of residue load. The average water absorption rate of bagasse fibres is estimated at 8% for the 2.5 mm fiber size and 12% for the 4 mm fiber size for all composites. Mechanical characterization by bending tests reveals a fragile behavior of the samples tested. Young’s modulus decreases when the load rate of fibrous residues increases regardless of the size of the fibers.

用AFLP分子标记技术鉴别甘蔗(Saccharum of ficinarum L.)品种

本研究采用AFLP技术 ,从 6 4对引物中筛选出 5对带型分布均匀、多态性丰富且分辨能力强的引物 ,对两个在生产上广泛推广应用的甘蔗品种的AFLP指纹图谱进行分析 ,计算了两品种间的遗传相似性和遗传距离。结果表明 ,5对引物在 2个甘蔗品种间均存在显著的差异 ,其中多态性位点占总扩增位点的 10 2 % ,区分率达 10 0 %。研究结果对生产上应用AFLP技术鉴定甘蔗品种的真伪性提供了依据。本文对甘蔗AFLP分析体系的关键技术以及银染技术的安全性和可靠性进行了讨论。

甘蔗细茎野生种(Saccharum spontaneum L.)的遗传多样性和系统演化研究

利用 RAPD技术 ,采用 2 5个随机引物对来自中国不同生态环境的 195份甘蔗细茎野生种材料进行了地理群体结构的遗传多样性研究。结果表明 :甘蔗细茎野生种的种内遗传变异较大 ,各地理类群的遗传分化明显 ,具有丰富的遗传多样性。基于分子聚类分析 ,推论中国甘蔗细茎野生种的起源演化方式为 :起源于云南 ,然后由云南→四川→贵州→广西→广东→海南→福建→江西 ;

两个割手密(Saccharum spontaneum L.)无性系对UV-B辐射响应的形态和生理差异

采用田间试验方法,研究了连续两年大田条件下,增强UV-B辐射(UV-B,280￣310nm,5.0kJ·m-2)对两个割手密(Saccharum spontaneum L.)无性系(Ⅰ91-48和Ⅱ91-81)形态和生理的影响,以及两个割手密无性系对UV-B辐射响应的种内差异。结果表明,两年中,在增强UV-B辐射下,Ⅰ91-485个生育时期的株高(除2004年成熟后期外)和分蘖数,以及分蘖期和伸长初期的茎径显著增加,而Ⅱ91-81株高在伸长初期和伸长后期,分蘖数在伸长初期、伸长后期、成熟初期和成熟后期(除2004年伸长后期外),以及茎径在分蘖期、伸长初期和伸长后期显著降低。两年的成熟后期,UV-B辐射导致Ⅰ91-48生物量、叶面积指数和节间长显著增加,Ⅱ91-81生物量、叶面积指数和节间长显著降低,而两个无性系节数无显著变化。两年中,UV-B辐射显著降低两个割手密无性系伸长后期和成熟初期叶展开角度。在UV-B辐射下,两年的5个生育时期中,Ⅰ91-48的类黄酮含量显著增加,丙二醛含量无显著变化,而Ⅱ91-81的类黄酮含量显著降低(除2003成熟后期外),丙二醛含量显著增加。总体上,UV-B辐射导致Ⅰ91-48叶绿素a、叶绿素b和叶绿素(a+b)含量在两年的分蘖期、伸长初期和伸长后期显著增加,而Ⅱ91-81叶绿素a、叶绿素b和叶绿素(a+b)含量在两年的分蘖期、伸长初期、伸长后期和成熟初期显著降低。割手密无性系Ⅰ91-48和Ⅱ91-81对UV-B辐射的响应具有种内差异。

割手密(Saccharum spontaneum L.)幼叶的组织培养

研究了从割手密幼叶诱导愈伤组织的条件,试验表明,培养基以MS附加2,4-D1～4mg/L为佳,所接种的幼叶以4cm以内为适宜的区段。不同材料在诱导愈伤组织和分化成苗方面都表现出明显的差异,说明云南的割手密在生理和遗传上存在着丰富的种内变异.

四川割手密(Saccharum spontancum L.)同工酶研究

以四川省的48个割手密为材料,采用垂直板状聚丙烯酰胺凝胶电泳进行3种同工酶分析。结果表明,在酯酶、过氧化物酶、谷氨酸草酰乙酸转氨酶中,以过氧化物酶分析效果最佳,可作为甘蔗野生资源种群划分的生化指标。根据过氧化物酶谱特征,供试样本出现19个类群,反映了四川割手密的多态性、遗传差异和地理分布的特点。

甘蔗的分类──Ⅱ．Saccharum L．属

Ｓａｃｃｈａｒｕｍ Ｌ．属的分类，经历了较大的变化。现在受到普遍接受的是Ｂｒａｎｄｅｓ（１９５８）的分类方法。他把ＳａｃｃｈａｒｕｍＬ．属分为６个种。本文讨论该属分类的历史衍变过程，各个种的形态特征、遗传特点、及对甘蔗育种的重要性。

Genetic Research and Utilization Reference of Saccharum spontaneum L.

[Objectives]The genetic characteristics of wild germplasm resources of Saccharum spontaneum L.were further investigated to provide a scientific theoretical basis for the improved hybridization of sugarcane varieties.[Methods]The first flowering date data of 112 wild resources were statistically analyzed by broad heritability for 3 consecutive years,and pollen quantity and yield were investigated to provide reference for hybridization.[Results]The broad heritability of the first flowering date of S.spontaneum was 0.079.The differences in the first flowering date between the S.spontaneum resources were in the range of 4-49 d,with an average of 13.65 d.The inter-annual changes within 20 d accounted for 90 %,and only 4.5% exceeded 30 d.The first flowering dates of some S.spontaneum resources were obviously postponed after being treated.[Conclusions]In the hybridization season,the hybridization plan must be adjusted according to the actual situation of the flowering period to make full use of it.

Optimization of protoplast isolation,transformation and its application in sugarcane (Saccharum spontaneum L.)

Sugarcane is a prominent source of sugar and ethanol production.Genetic analysis for trait improvement of sugarcane is greatly hindered by its complex genome,long breeding cycle,and recalcitrance to genetic transformation.The protoplast-based transient transformation system is a versatile and convenient tool for in vivo functional gene analysis;however,quick and effective transformation systems are still lacking for sugarcane.Here,we developed an efficient protoplast-based transformation system by optimizing conditions of protoplasts isolation and PEG-mediated transformation in S.spontaneum.The yield of viable protoplasts was approximately 1.26×107 per gram of leaf material,and the transformation efficiency of 80.19%could be achieved under the optimized condition.Furthermore,using this approach,the nuclear localization of an ABI5-like bZIPs transcription factor was validated,and the promoter activity of several putative DNase I hypersensitive sites(DHSs)was assessed.The results indicated this system can be conveniently applied to protein subcellular localization and promoter activity assays.A highly efficient S.spontaneum mesophyll cell protoplast isolation and transient transformation method was developed,and it shall be suitable for in vivo functional gene analysis in sugarcane.

Energy Productivity Potential Assessment of Saccharum spontaneum L. Germplasm Resources

[Objective]The energy productivity potential of Saccharum spontaneum L.germplasm resources as energy plants was discussed in this paper.[Method]The energy productivity potential of Saccharum spontaneum L.germplasm resources collected from the wild was assessed from the aspects of cellulose,semicellulose,lignin and crude ash contents and caloric value.[Result]The total content of cellulose and semicellulose ranged from 41.75%to 69.13%,the lignin content ranged from 2.16%to 11.75%,the crude ash content ranged from 4.79%to 9.34%and the caloric value ranged from 16.00 to 17.69 MJ/kg among the 30 Saccharum spontaneum L.germplasms.Compared to other plants,Saccharum spontaneum L.had higher contents of cellulose and semicellulose,a higher caloric value but lower contents of lignin and crude ash.[Conclusion]From the perspective of producing fuel ethanol with lignocellulose,Saccharum spontaneum L.has a great potential for development as an energy plant.

Nutritional Value Evaluation of Saccharum spontaneum L. Germplasm Resources

The contents of dry matter, crude fat, crude protein, neutral detergent fiber, acid detergent fiber, crude ash, Ca and P in 43 Saccharum spontaneum L.germplasms during the vegetative period were determined. Among them, the crude protein, crude fat, neutral detergent fiber, acid detergent fiber, P and Ca contents were treated as the judging indicators. The nutritional value evaluation was carried out with the analytic hierarchy process(AHP). The results showed A49(Guangxi)had the highest nutritional value. Its dry matter, crude protein, crude fat, crude ash,neutral detergent fiber, acid detergent fiber, Ca and P contents were 32.35%,7.20%, 4.06%, 8.07%, 75.81%, 50.72%, 0.23% and 0.16% respectively. While A3(Guangdong) had the lowest nutritional value. Its dry matter, crude protein, crude fat, crude ash, neutral detergent fiber, acid detergent fiber, Ca and P contents were32.24%, 4.62%, 0.51%, 6.38%, 46.40%, 40.02%, 0.15% and 0.09% respectively.The crude protein content differed significantly among different germplasms. A48 had the highest crude protein content(9.11%), and A14 had the lowest crude protein content(3.72%). Based on the evaluation results, the 43 Saccharum spontaneum L.germplasms were divided into 3 groups: high-nutritional value type, moderate-nutritional value type and low-nutritional value type. We hoped to provide a theoretical reference for the application of Saccharum spontaneum L. as a forage grass.

Study on Saccharification of Saccharum spontaneum L. by Sulphuric Acid Hydrolysis

In order to improve the saccharification rate by acid hydrolysis in the technique for production of ethanol from biomass,in this study,Saccharum spontaneum L. was used as the experimental material to investigate the rules and technological conditions for two-step acid-hydrolysis saccharification by single factor experiment and orthogonal experiment. According to the results,concentrated sulfuric acid hydrolysis was the first step,with the liquid-solid ratio of 42∶ 1,sulfuric acid concentration of 70%,hydrolysis time of 20 min,and hydrolysis temperature of 55 ℃; dilute sulphuric acid hydrolysis was the second step,with the liquidsolid ratio of 115∶ 1,sulfuric acid concentration of 5. 5%,hydrolysis time of 155 min,and hydrolysis temperature of 100 ℃. In two-step acid hydrolysis process,the yield of reducing sugar was 48. 78%. The results indicated that two-step acid-hydrolysis saccharification required mild conditions,simple operation and low cost,and led to high reducing sugar yield,exhibiting a broad application prospect.

甘蔗NBS-LRR类抗病基因同源序列的分离与鉴定

根据已知植物(拟南芥、烟草和亚麻)抗病基因(RGAs)保守序列设计简并引物,从甘蔗高抗黑穗病品种NCo376的基因组DNA和cDNA中扩增出11条抗病基因同源序列,其中5条来自基因组DNA(EF059973、EF059974、EF059975、EF059976和EF059977),6条来自cDNA(EF155648、EF155649、EF155650、EF155651、EF155652和EF155653)。序列分析表明,这些RGAs均含有典型的NBS-LRR类抗病基因所拥有的保守结构域P-loop、Kinase-2a、Kinase-3a和疏水结构域。聚类分析表明,11条RGA同RPS2和XA1聚为一类,而N和L6则单独聚为一类。所有11条抗病基因同源序列中,kinase-2(LLVLDDVW/D)最后一个氨基酸皆为色氨酸。定量PCR分析表明,编号为EF059974的PIC基因的表达不仅受黑穗病菌胁迫的影响,而且受水杨酸的诱导和过氧化氢的抑制,也具有抗病基因组织特异性和组成型表达特性。

割手密SnRK2基因家族全基因组鉴定及干旱胁迫表达分析

【目的】探究割手密SnRK2基因家族成员在干旱胁迫中的调控机制,为抗旱性甘蔗品种的选育提供侯选基因。【方法】以全基因组数据为基础从割手密中鉴定SnRK2基因,并对其进行生物信息学分析和干旱胁迫下的表达分析。【结果】在割手密基因组中共鉴定出11个SnRK2基因家族成员,命名为SsSnRK2.1-SsSnRK2.11,且这些基因不均匀地分布于8条染色体上。SnRK2蛋白的氨基酸残基数为227~580,分子质量为25683.53~64695.8 kD,等电点为4.62~8.94,且均为亲水性蛋白。系统发育树可将其分为3个亚组,且同亚组中的保守基序基本相似,外显子数量以7~9个为主。SsSnRK2基因家族成员的启动子中含有多种激素类和逆境胁迫响应类的作用元件。割手密SsSnRK2基因家族成员的表达具有组织特异性。所有的SsSnRK2基因均能不同程度地响应干旱胁迫。【结论】割手密SnRK2基因家族在响应干旱胁迫过程中发挥重要作用,可为割手密的抗逆性研究提供一定的理论依据。

30份割手密种质资源苗期抗旱性综合评价

【目的】依据生理生化指标综合评价不同基因型割手密种质资源的苗期抗旱性,为甘蔗抗旱性育种的亲本选育提供参考。【方法】以30份割手密野生种质为试验材料进行盆栽试验,测定割手密苗期干旱胁迫处理前后的6项生理生化指标,采用主成分分析、模糊隶属函数分析和聚类分析对割手密的抗旱性进行综合评价。【结果】与正常供水相比,干旱胁迫下各割手密种质材料的过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、过氧化物酶(POD)活性增强,丙二醛(MDA)、可溶性糖(SS)、脯氨酸(Pro)含量增加,但各指标的变化程度因材料不同而存在一定差异。相关性分析结果表明,在干旱胁迫下,不同割手密种质材料叶片的过氧化氢酶活性和脯氨酸含量之间呈显著正相关(P<0.05),超氧化物歧化酶活性和脯氨酸之间呈极显著正相关(P<0.01);过氧化物酶活性和超氧化物歧化酶活性之间呈显著负相关;过氧化氢酶活性与丙二醛含量之间也呈显著负相关,其他指标之间无显著相关性(P>0.05)。主成分分析表明,SOD活性、POD活性、CAT活性和Pro含量、SS含量、MDA含量可作为割手密抗旱性评价的重要指标。通过模糊隶属函数分析和聚类分析可将30份割手密种质材料划分为4类,其中高度抗旱材料1份(90⁃53),中高度抗旱材料2份,中度抗旱材料14份,低度抗旱材料13份。【结论】CAT活性、SOD活性、POD活性、MDA含量、SS含量、Pro含量可作为割手密苗期的抗旱性评价指标。30份参试割手密种质中,90⁃53的抗旱性最强,2015⁃22、88⁃301次之,2015⁃104的抗旱性最弱。

甘蔗耐盐生理研究 IV.NaCl胁迫对甘蔗多胺代谢影响

应用盆栽试验详细研究了NaCl胁迫下甘蔗叶片多胺代谢的变化.结果表明:随NaCl胁迫强度的增加,ADC、ODC和PAO活性以及多胺含量都显著或极显著提高,而且它们之间均呈极显著正相关,说明NaCl胁迫下诱发的多胺含量提高与ADC、ODC和PAO关系密切.而且ADC、PAO活性和多胺相对值的增加与品种耐盐性表现一致,耐盐性强的品种桂糖11和NCo310 ADC活性提高幅度高于不耐盐品种粤糖57—423,导致多胺含量提高;而PAO活性不耐盐品种增加高于耐盐品种.然而ODC活性变化似乎与品种耐盐性关系不甚密切,仅为NaCl胁迫的适应性反应.最后本文提出了NaCl胁迫下甘蔗的多胺代谢及其可能的适应机制.