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.

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.

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。

Evaluation of Soil Quality Indicators in Sugarcane Management in Sandy Loam Soil

An important factor for the sustainability of soils highly susceptible to degradation is the use of monitoring tools that promptly and realistically reflect changes imposed on soil by different cropping systems. To select soil quality indicator variables in sugarcane (Saccharum officinarum L.) production areas that fulfill the criteria of sensitivity to management practices and between-season consistency in the management discrimination, ten composite soil samples (0–10 cm) were collected in July 2005 (rainy season) and again in March 2006 (dry season) from areas under cultivation of organic sugarcane (OS), green sugarcane (GS), burned sugarcane (BS) and from an adjacent native forest (NF) area at Usina Triunfo, Boca da Mata, Alagoas, Brazil. Microbial biomass-C (MBC), total organic C (TOC), soil enzyme activity expressed as the rate of fluorescein diacetate (FDA) hydrolysis, mean weight diameter of water-stable soil aggregates (MWD), and percentage of water-stable macroaggregates (PWSA) were analyzed. Although MBC and TOC were higher in NF than in the cultivated areas, no differences were observed in these C pools between the three sugarcane systems. The response of FDA to the site management was dependent on the sampling time. In the rainy period, the activity followed the order: NF > OS > GS > BS, whereas in the dry season, only NF differed from the other treatments. Irrespective of the sampling time, MWD and PWSA decreased in the order NF > OS = GS > BS. The variables MWD and PWSA are quite sensitive for discriminating between site management histories regardless the sampling season.

<i>In Vitro</i>Priming of Sugarcane Varieties (RB966928 and RB867515)

Sugarcane, as a glycophyte, shows sensitivity to saline soils at various stages of its growth. The aim of this study was to evaluate the in vitro priming response in two sugarcane varieties (RB966928 and RB867515). Micropropagated plants, from meristems, received priming treatments by adding the salts (NaCl and KCl) in different concentrations (0.0;12.5;25.0 and 50.0 mM), in the MS medium. Subsequently, the plants were cultivated in rooting medium without addition of salts, acclimatized and submitted to gradient ex vitro saline stress with 20 → 40 and 60 mM, of each salt, for 30 days. The analyzed variables were dry matter of shoot and root, number of tillers and estimation of chlorophyll content. The experiment was carried out in a 2 × 2 × 4 factorial arrangement, in a completely randomized experimental design. Twenty replicates were used throughout the experiment. Data were submitted to analysis of variance and regression and the means were compared by Tukey’s test, at a 5% probability level. The priming treatments presented a significant effect, with triple interaction, in the chlorophyll index. In the treatment with NaCl, the variety RB966928 showed an increase in the chlorophyll index with the increase of treatment levels, up to an optimal limit of 31.47 mM. On the other hand, the variety RB867515 showed decreasing in chlorophyll index. In contrast, in KCl treatment, the variety RB867515 presented the increase at the chlorophyll index with the maximum point of 25 mM. For the variables, shoot dry matter (SDM) and root dry matter (RDM) there was a significant difference (p 0.05 and p 0.01, respectively) only between the varieties. The variety RB966928 presented higher SDM and RDM in relation to the variety RB867515. Studies are recommended with increasing the duration of the priming treatments and more detailed study of the culture throughout its productive cycle.

Infestation of Diatraea saccharalis(Fabr.)during Five Consecutive Cycles of Sugarcane Crop Succeeding Leguminous Crops

The area cultivated with sugarcane in Brazil is expanding, and increasingly using legumes cover crop in new deployed areas and in those which were reformed. Long-term trials were carried out in Piracicaba, Sao Paulo, Brazil, to evaluate the effect of leguminous green manure on the natural infestation of Diatraea saccharalis (Fabr.) during five cropping cycles of sugarcane “IAC 87-3396”. This study was carried out from October 1999 to October 2005 in Piracicaba, Sao Paulo, Brazil, at coordinates 22°42''S and 47°38''W, 560 m above sea level, in soil classified as Typic Paleudult. Green manure crops were assigned to five treatments: peanuts (Arachis hypogaea L.) (cultivar “IAC-Tatu” and “IAC-Caiapó”), sunn hemp (Crotalaria juncea L.) (cultivar “IAC 1”) and velvet-bean [Mucuna aterrima (Piper & Tracy) Holland] (cultivar “common”). A treatment without any green manure or weed was used as the control. The previous sugarcane crop was destroyed and the soil was prepared in a conventional way (using plow and harrow) before the leguminous crops were sowed. Thirty days after the sowing, weeds were removed and, 120 days later, the crops were cut and its biomass was deposited on the soil surface without incorporation. Experimental plots comprised five rows of sugarcane, each one measuring 10 m long and spaced 1.40 m apart. The sugarcane was harvested 18 months after biomass deposit, on October 25th, 2001 and its stalks were collected at intervals of approximately 12 months in: 07/09/2002, 08/01/2003, 11/07/2004 and 10/06/2005. At harvest, the masses were determined from a sample of sugarcane, cut from three rows (2 m long) in the center of the plot. The losses caused by the sugarcane borer were estimated by the intensity of the infestation by randomly observing and collecting sugarcane stems from each plot. A great reduction in the number of stalks and in the yield was noted proportionally to the intensity of the infestation of the borer in the sugarcane crop grown after the velvet-bean. The sugarcane-velvet-bean rotation should be regarded considering the intensity of sugarcane borer infestation. No influence of the other green manure crops was observed on the intensity of the infestations. However, the harvest seasons of the sugarcane seemed to influence the borer infestation.

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。

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。

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。

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.

NaCl胁迫对甘蔗生长影响的机理研究

４种ＮａＣｌ浓度（０．２５％、０．５０％、０．７５％和１．００％）处理，显著降低了甘蔗株高、茎径、地上部盆干重和蔗汁蔗糖份；耐盐性强的品种如桂糖１１和Ｎｃｏ３１０的降幅小于不耐盐品种（粤糖５７－４２３），而且这些变化与ＮａＣｌ胁迫引发的叶绿素含量下降和蛋白酶活性上升有关。最后本文还提出了ＮａＣｌ胁迫抑制甘蔗生长的可能机理，认为ＮａＣｌ胁迫下产生的Ｎａ＋离子累积毒害和叶片水份状况恶化是盐害的原初效应，而非次级效应。

广西割手密核心种质构建与关联分析

为发掘甘蔗育种优异野生基因资源,以来自广西的333份割手密为材料,应用12对SSR引物的分子标记数据和28个表型性状资料构建广西割手密核心种质,并进行关联分析。关联分析结果显示,广西割手密茎径、节间长度、曝光前颜色、脱叶性和毛群共5个表型性状与8个位点显著相关;茎径与叶长、叶宽、节间长度、节数和锤度5个性状均表现出极显著相关;株高、茎径、节间长度、节数4个表型性状之间呈现显著或极显著的正相关;锤度与茎径和节间长度均表现为极显著的负相关。核心种质抽样按照总资源比例的0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9进行梯度筛选,当抽样比例达到30%以上时,即可包含100%等位基因覆盖度。遗传多样性评价和主成分分析结果显示,构建割手密核心种质所筛选材料具有丰富的遗传多样性,且基于核心种质绘制的主成分图与原始种质的分布图趋势相吻合。结果表明,根据30%的抽样比例筛选出99个割手密样本构建核心种质,遗传多样性评价和主成分分析所构建的核心种质具有较好的代表性。

多元统计分析与投影寻踪法对甘蔗品质性状综合评价

本研究以新台糖22号及其7个杂交后代甘蔗新品种(系)为研究对象,利用多样性分析与投影寻踪法对甘蔗蔗糖分、纤维分等11个品质指标进行综合评价分析。结果表明,甘蔗蔗糖分与蔗汁直接转光度、蔗汁转化转光度呈极显著正相关,甘蔗纤维分与出汁率、甘蔗产糖量、蔗渣含水量呈极显著负相关,与蔗渣转光度呈显著正相关。通径分析结果表明,甘蔗糖分相关指标(蔗汁锤度、蔗汁蔗糖分、重力纯度、蔗汁直接转光度及煮液锤度等)对甘蔗蔗糖分具有直接正效应,蔗汁糖分及蔗渣煮液等相关指标(蔗汁锤度、重力纯度、蔗汁转化转光度、煮液转光度)对甘蔗纤维分具有直接正效应。对13个品质指标进行主成分分析,将其归为3个主成分,其特征值累积贡献率达85.71%。基于主成分分析及投影寻踪分类(PPC)分析,对不同甘蔗品质指标进行综合评价发现,桂糖09-439的综合评分值最高。聚类分析可将甘蔗品种(系)划分为三大类群,河糖11-157为第Ⅰ类群,为高糖高纤维分品种;桂糖09-439为第Ⅱ类群,为早熟高糖品种。本研究运用多元统计分析与投影寻踪法,都筛选出同一高糖甘蔗种质资源品种桂糖09-439、河糖11-157为高糖高纤维分品种,同时发现投影寻踪分类法可作为初筛大量甘蔗种质资源的途径之一。

Skin Anaphylaxis Test of Purified Protein HAS1 in Cavies

The antifungal protein HAS1 is a new antifungal protein isolated from the genome of Bacillus subtilis HAS,which has a good inhibitory effect on various pathogenic fungi in sugarcane.This study aimed at evaluating the immunological transfer reaction of the test sample through repeated skin contact by observing whether the cavy skin repeatedly exposed to purified protein HAS1 suffers from allergic reaction and how strong is the allergic reaction.The results showed that the test group and the vehicle control group exhibited no allergic reaction in the skin immediately and at 24,48 and 72 h and had an mean reaction score and a sensitization rate both of 0,so the results of the naked eye observation were both nonallergenic to cavy skin;and in the positive control group,the cavies were observed to be highly sensitized immediately after removing the drug,mildly sensitized at 24 h,and not sensitized at 48 and 72 h,which meant the skin allergy was alleviated with the observation time.It is suggested that the purified protein HAS1 is negative for cavy skin anaphylaxis test (nonallergenic to cavy skin),which provides an experimental basis for further utilization of the protein and its coding gene.

美国引进甘蔗热带种的ARMS PCR真实性鉴定

甘蔗遗传基础狭窄已经极大地阻碍了其育种进程,挖掘优异种质资源、创新亲本材料将成为未来甘蔗育种的重要方向之一。利用四引物扩增受阻突变PCR体系(ARMS PCR)对国家甘蔗种质资源圃保存的美国引进的24份疑似热带种材料进行真实性鉴定。PCR鉴定结果表明,供试材料中有‘Green German’、‘Black Fiji’、‘Chittan’、‘IN 84-072’和‘NG 21-002’这5份材料扩增带型与热带种‘拔地拉’(Badila)一致,仅出现428 bp的共有条带和278 bp的热带种特有条带,属于热带种带型;而其它19份材料同时具有278 bp的热带种特有条带和203 bp的割手密特有条带,属于热带种与割手密的杂交种带型。经过上述分子鉴定得出结论,24份美国引进的甘蔗种质中有‘Green German’、‘Black Fiji’、‘Chittan’、‘IN 84-072’和‘NG 21-002’这5份材料为热带种材料,建议后续加强其农艺性状和抗性调查,并作为甘蔗热带种新血缘亲本进行杂交利用。

广西甘蔗生育期内气候资源空间特征及对气候变化的响应

利用1961—2020年广西91个国家气象观测站逐日气象数据,对甘蔗(Saccharum officinarum L.)苗期(P1)、分蘖期(P2)、茎伸长期(P3)和工艺成熟期(P4)气候资源空间分布特征进行分析,同时采用线性趋势分析方法探讨甘蔗生育期内气候资源对气候变化的响应。结果表明,在空间分布上,平均气温在苗期西高东低,在茎伸长期和工艺成熟期南高北低;日照时数在苗期西高东低,在茎伸长期东高西低,在分蘖期和工艺成熟期南高北低;降水量在苗期和分蘖期东、东北高西南低,在茎伸长期西、西南高东北低,在工艺成熟期桂中地区高、其他区域低。气候变化背景下,甘蔗生育期内平均气温显著上升,在桂北甘蔗工艺成熟期上升最快;日照时数显著下降,在茎伸长期时最明显;降水量在桂东北甘蔗分蘖期呈部分显著上升趋势。

甘蔗不同品种光合特性比较及其聚类分析

利用CI-301PS光合作用测定系统测定152个甘蔗品种资源的光合参数。结果表明,甘蔗品种间光合性状存在较大差异。进一步对152个甘蔗品种资源的光合性状进行聚类和判别分析,将152个品种分为7大类,在聚类分析基础上用判别分析选出对甘蔗光合性状数值分类有显著影响的3个光合参数,建立了7个判别能力较高的判别模型。

甘蔗不同叶位叶片形态与冠层特征的关系

利用CI-100数字植物冠层分析仪和CI-203叶面积分析仪测定了 4个甘蔗品种不同叶位层的冠层参数和相应叶位叶片的形态特征,结果表明:不同基因型不同叶位层间叶片形态、冠层空间结构和冠层辐射特征存在显著差异,不同叶位层叶面积指数、叶簇倾角和叶分布的变化主要由冠层内不同叶位叶片叶宽的变化引起,而不同叶位层消光系数的变化主要与叶簇倾角和叶分布有关;散射光透过系数的变化主要与叶面积指数、叶簇倾角和叶分布有关,直射光透过系数的变化主要与叶分布、叶宽、长宽比有关,光合有效辐射的变化与叶面积指数、叶面积、叶宽、长宽比有关.研究结果同时表明,叶片形态与冠层辐射特征存在显著的典型相关关系. 图 2表 4参

能源甘蔗不同叶位叶片形态、光合气体交换及其与产量关系

利用CI-203叶面积分析仪和CI-301CO2气体分析仪分析4个能源甘蔗品种分蘖期和伸长初期不同叶位层叶片的形态特征和光合气体交换特征参数.结果表明,不同基因型不同叶位层间叶片形态、光合气体交换特征参数存在显著差异.分蘖期和伸长初期蔗叶的叶长、叶宽、叶面积、长宽比等叶片形态参数和净光合速率、蒸腾速率、气孔导度和水分利用率等光合气体交换特征参数均表现为随着叶位的下降,自上而下逐渐变小.逐步回归分析结果表明,分蘖期第3叶的叶宽和水分利用率及第5叶的叶宽对产量形成影响较大;而伸长初期第1叶和第3叶的叶面积对产量形成影响较大.