割手密遗传作图最适分离群体的选择

[目的]筛选适合用于构建割手密遗传连锁图谱的分离群体,为后续的遗传连锁图谱构建和基因定位及遗传育种提供理论依据.[方法]基于“双假测交“理论,利用DNA水平上遗传差异较大且蔗糖分和分蘖率差异极显著的不同割手密无性系进行杂交获得3个不同杂交组合,对3个备选作图群体的分蘖率和蔗糖分进行变异、正态分布拟合分析及分子多态性分析.[结果]组合3（GXS85-12×GXS87-16）和组合13（云割①×田林）的分蘖率呈偏态分布,而组合5（GXS85-30×GXS87-16）的分蘖率呈近似正态分布;组合3和组合5的蔗糖分呈偏态分布,而组合13的蔗糖分呈近似正态分布.在分子多态性水平上,组合5亲本的遗传相似系数略高于组合3和组合13,而其后代平均遗传相似系数略低,但变幅较高.[结论]组合5（GXS85-30×GXS87-16,群体大小为245株）的分蘖率、蔗糖分遗传性相对稳定,群体来源比较广泛,分子多态性丰富,遗传多样性高,可作为分离群体用于构建遗传图谱.     

甘蔗与斑茅割手密复合体的回交后代真实性鉴定及染色体遗传分析

【目的】掌握斑茅割手密复合体(简称斑割复合体)在杂交利用过程中的染色体遗传规律,为斑割复合体适宜回交利用代数提供参考依据。【方法】选用4对引物对甘蔗与斑茅割手密复合体的回交后代进行SSR标记鉴定,并利用甘蔗根尖细胞酶解去壁低渗法对真实杂种进行染色体数目观察。【结果】供试的28个杂交后代材料(15个GT05-164与GXASF108-2-28杂交BC1代,13个GT42与GXASBC112-A6-25杂交BC2代)均为真实杂种。选择在2对SSR引物中均扩增出父本特征带的后代(5个BC1代和7个BC2代)进行染色体观察,结果发现,5个BC1代的染色体数目介于95~98条,其染色体按“n+n”方式传递;7个BC2代的染色体数目介于94~101条,其染色体也按“n+n”方式传递。【结论】甘蔗与斑割复合体杂交BC1和BC2代中染色体传递方式均为“n+n”,母本甘蔗的染色体未加倍,高贵化育种进程减慢,可能需要更高的杂交代数才能获得聚集斑茅和割手密血缘的优良品种(系)。     

Photochemical damage and comparative performance of superoxide dismutase and ascorbate peroxidase in sugarcane leaves exposed to paraquat-induced oxidative stress

The physiological responses of sugarcane (Saccharum officinarum L.) to oxidative stress induced by methyl viologen (paraquat) were examined with respect to photochemical activity, chlorophyll content, lipid peroxidation and superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities. Thirty-day-old sugarcane plants were sprayed with 0, 2, 4, 6 and 8 mM methyl viologen (MV). Chlorophyll fluorescence was measured after 18 h and biochemical analyses were performed after 24 and 48 h. Concentrations of MV above 2 mM caused significant damage to photosystem II (PSII) activity. Potential and effective quantum efficiency of PSII and apparent electron transport rate were greatly reduced or practically abolished. Both chlorophyll and soluble protein contents steadily decreased with MV concentrations above 2 mM after 24 h of exposure, which became more pronounced after 48 h, achieving a 3-fold decrease. Insoluble protein contents were little affected by MV. Oxidative stress induced by MV was evidenced by increases in lipid peroxidation. Specific activity of SOD increased, even after 48 h of exposure to the highest concentrations of MV, but total activity on a fresh weight basis did not change significantly. Nondenaturing PAGE assayed with H₂O₂ and KCN showed that treatment with MV did not change Cu/Zn-SOD and Mn-SOD isoform activities. In contrast, APX specific activity increased at 2 mM MV but then dropped at higher doses. Oxidative damage induced by MV was inversely related to APX activity. It is suggested that the major MV-induced oxidative damages in sugarcane leaves were related to excess H₂O₂, probably in chloroplasts, caused by an imbalance between SOD and APX activities, in which APX was a limiting step. Reduced photochemical activity allowed the early detection of the ensuing oxidative stress.     

Modulation of carbohydrate metabolism and chloroplast structure in sugarcane leaves which were infected by Sugarcane Yellow Leaf Virus (SCYLV)

Non-symptomatic sugarcane plants infected with Sugarcane Yellow Leaf Virus showed starch in mesophyll and bundle sheath cells. In situ-hybridization of mRNAs of sucrose-phosphate phosphatase and ADP-glucose pyrophosphorylase revealed that infected leaves contained SPPase and AGPase in mesophyll cells, Kranz cells and bundle sheath cells. In contrast virus-free leaves contained SPPase only in Kranz cells and AGPase only in bundle sheath cells. Infected leaves exhibited ultrastructural changes in Kranz cell chloroplasts and a shift of the chlorophyll a/b ratio. No obstruction of plasmodesmata was observed. The results indicate that SCYLV-infected plants, even when visually non-symptomatic, underwent strong metabolic and ultrastructural changes.     

中国本土割手密血缘创新亲本材料的利用潜力分析

 以云割F1,F2,崖城71-374（F2）及其后代与国内外常用亲本配成的30个杂交组合为研究材料,分析其有性世代主要性状的遗传效应。结果表明：母本、父本及选配方式的不同后代的主要性状均表现极显著差异（P<001）,平均遗传力依次为：组合（896%）＞父本（764%）＞母本（706%）;后代主要性状的遗传力大小顺序为锤度＞有效茎＞茎径＞株高＞蔗产量＞糖产量;云割F197-105,云割F197-147,云割F203-7,云割F203-78,云割F180-189,云瑞05-458,云割F203-72等7个材料做父本的一般配合力优于优良亲本崖城71-374,可作为优良创新种质、亲本材料重点利用;云蔗99-155,云割F203-315,云蔗89-7,桂糖92-66,内江00-118可作为高糖亲本利用;德蔗93-88×云割F203-7,CP65-357×云割F180-189,粤糖93-159×云割F180-189可作为高产、高糖优良创新组合重点利用;CP65-357×云割F197-105,粤糖93-159×云割F197-147,云瑞05-344×云割F197-105,Q141×云割F197-105,云瑞05-189×云割F203-78,云瑞05-344×云瑞05-458,农林03-1×云割F197-105等可作为高生物量能源甘蔗育种优良创新组合重点利用。     

斑割复合体及其杂交后代的生物量及根系性状

本研究利用前期获得的含广西斑茅和广西割手密血缘的甘蔗育种新材料及其亲本共11份无性系进行桶栽试验,分析了参试材料植株地上部生物量和根系的主要相关性状在斑茅割手密复合体杂交利用过程各世代的变化。结果表明:参试的斑茅割手密复合体及其与甘蔗杂交、回交获得的F1代材料以及BC1代在株高、茎径、单茎重和叶面积上都表现比其亲本甘蔗有更大的优势;根系重量和活力都略有增加;地上部分生物产量上均表现出超亲效应,其超亲效应与绿叶面积的超亲表现相关性显著。     

Genotypic variation of resistance to the spotted stalk borer Chilo sacchariphagus (Bojer) in sugarcane: evidence of two distinct resistance mechanisms

We carried out an assessment of the resistance status to the spotted stalk borer Chilo sacchariphagus in a sample of 32 international sugarcane clones in Reunion. Our objectives were to characterize the genotypic variability of resistance and to characterize the role of leaf and stalk resistance in the global resistance. A significant genotypic variability in the % exited tunnels and in the leaf feeding injuries confirmed the existence of two resistance mechanisms located respectively in the leaves and in the stalk. Leaf resistance and stalk resistance exhibited genotypic and phenotypic correlation with stalk tunnelling injuries. No phenotypic or genotypic correlations were observed between leaf resistance and stalk resistance. Both resistances contributed independently to the global resistance quantified by the stalk tunnelling. The resistance status of the two main cultivars cultivated in Reunion was assessed within our germplasm sample: 'R 570' was one of the most resistant clones and 'R 579' proved to be moderately susceptible.     

Genetic diversity associated with in vitro and conventional bud propagation of Saccharum varieties using RAPD analysis

Polymorphisms in the genomic DNA of eight varieties maintained by conventional bud propagation (via rhizomes) and by in vitro shoot tip cultures were detected by RAPD analysis of sugarcane varieties. The study estimated the genetic diversity induced after in vitro multiplication of these varieties. Higher (28.9%) and lower (12%) numbers of polymorphic bands were detected in plants propagated via rhizomes; the genetic similarity estimate varying from 0.63 to 0.80. Plants of SP90‐3723 and SP91‐1049, or RB85‐5113 and SP90‐3723, varieties involving greater genetic distances may be indicated as progenitors in breeding programmes. In vitro multiplication of RB86‐7515, RB85‐5113, RB83‐5054 and SP86‐42 varieties increases genetic variability, while in vitro multiplication of SP91‐1049, SP90‐1638, RB92‐8064 and SP90‐3723 leads to genetic similarity. Results show that the RAPD technique is an effective tool for detecting polymorphism in sugarcane clones and it allows quick collection of the necessary information (more genetically divergent plant varieties) to guide new crossing in sugarcane breeding programmes.     

Effects of Lactobacillus buchneri isolated from tropical maize silage on fermentation and aerobic stability of maize and sugarcane silages

This study was aimed to perform a screening of Lactobacillus buchneri strains from maize silage and use them as inoculant in maize and sugarcane silages. In all, 151 lactic acid bacteria (LAB) strains were isolated from whole‐plant maize silage, and their identification was based on the sequence analysis of 16S rDNA. In total, 15 strains were categorized to the L. buchneri group and eight of these were selected based on growth rate and fermentation pattern. The selected strains were evaluated on fermentation and aerobic stability of maize and sugarcane silages. For maize, the inoculated silages had lower pH and higher LAB population, but lower acetic acid concentration in comparison with the untreated control silage. For sugarcane silage, the strains 56.1, 56.4 and 40788 resulted in highest dry‐matter (DM) content and lowest DM losses. However, only the strain 40788 showed lowest counts of yeasts and moulds. Sugarcane silages inoculated with the strains 56.9, 56.26 and the untreated control silage showed highest concentrations of lactic acid and ethanol, besides the great DM losses. Even so, for both crops, the aerobic stability was not affected by inoculation. After air exposure, all silages increased temperature and had high population of yeast and moulds. Nevertheless, the strains 56.1 and 56.4 are promising for use as a silage inoculant.     

Distinguishing sporulating and nonsporulating lesions as a method for evaluating the resistance of sugarcane genotypes to orange rust

Sugarcane breeding programmes rank the resistance of genotypes to Puccinia kuehnii, causal agent of orange rust, according to levels of disease severity. However, during the screening stages, this method of assessment can lead to precipitous elimination of genotypes with promising agronomic traits but showing mild symptoms of rust such as flecks or lesions that do not produce spores. This study aimed to propose a new method to classify the resistance of sugarcane genotypes to orange rust by counting sporulating lesions. Five sugarcane varieties with different levels of resistance to P. kuehnii were inoculated with two pathogen populations under controlled conditions. The disease severity (SEV), total number of lesions (TNL), and total number of sporulating lesions (TNSL) were evaluated in a 20 cm leaf fragment from the most diseased leaf. The TNL and TNSL evaluations were performed at 11, 16 and 21 days after inoculation (DAI) and SEV at 21 DAI. The thresholds of 80% and 8% of sporulating lesions (SL) separated susceptible from the intermediate varieties and intermediate from the resistant ones, respectively. It is proposed that the method of counting sporulating lesions be used in screening genotypes for resistance to P. kuehnii in sugarcane breeding programmes.