Molecular Markers for Sweet Sorghum Based on Microarray Expression Data

Using an Affymetrix sugarcane genechip, we previously identified 154 genes differentially expressed between grain and sweet sorghum. Although many of these genes have functions related to sugar and cell wall metabolism, dissection of the trait requires genetic analysis. Therefore, it would be advantageous to use microarray data for generation of genetic markers, shown in other species as single-feature polymorphisms (SFPs). As a test case, we used the GeSNP software to screen for SFPs between grain and sweet sorghum. Based on this screen, out of 58 candidate genes, 30 had single-nucleotide polymorphisms (SNPs) from which 19 had validated SFPs. The degree of nucleotide polymorphism found between grain and sweet sorghum was in the order of one SNP per 248 base pairs, with chromosome 8 being highly polymorphic. Indeed, molecular markers could be developed for a third of the candidate genes, giving us a high rate of return by this method.     

高梁种质资源的创新和利用

高梁品种遗传改良取得的成果很大程度上取决于占有的高梁种质资源的数量,以及对其主要性状的鉴定、创新和利用。在高粱育种史上,品种改良的突破性进展,往往都是由于找到并利用了具有关键基因的种质。目前,已创造和选育出7种不同细胞质雄性不育系,其育性反应各不一样。除A．细胞质不育系组配的杂交种得到广泛应用外,A：细胞质不育系的杂交种在生产上应用,A,细胞质不育系在甜高粱杂交种上应用。籽粒优质、茎秆高糖、低氰氢酸、抗病虫、抗杂草、抗干旱等抗性种质资源也得到了创新利用。     

Alternate crop and weed host plant oviposition preferences by the Mexican rice borer (Lepidoptera: Crambidae)

The Mexican rice borer, Eoreuma loftini (Dyar), is the key pest of sugarcane, Saccharum hybrids, in south Texas, having largely displaced the sugarcane borer, Diatraea saccharalis (F.), and it is moving into rice- and sugarcane-growing areas of east Texas and Louisiana. While a number of alternative weed and crop hosts have been reported, the extent to which they might support Mexican rice borer populations is unknown. This study involved choice assays that compared oviposition preference for and larval infestations of five mature graminaceous weed species. Levels of infestation between sugarcane and corn, Zea mays L., crop hosts and between corn and sorghum, Sorghum bicolor (L.) Moench, were also assessed. We determined that the average number of larval entry holes in sudangrass stems was ≥2.5-fold more than for any of the other four weed host plants, that corn had ≥5.9-fold more larval entry holes than sorghum and ≥8.2-fold more than sugarcane. Greater oviposition and infestation of one non-crop host over another was not related to numbers of stems per plant, but was associated with the greater stem diameter and abundance of dry leaf tissue found in Sudangrass, Sorghum bicolor (L.) Moench ssp. drummondi (Nees ex Steud.) de Wet & Harlan, johnsongrass, S. halepense (L.) and barnyardgrass, Echinochloa crus-galli (L.) P. Beauv.; relative to the other weed species in this study. In terms of the crop plants, stalk diameter and quantity of dry leaf tissue were not associated with numbers of eggs or larval entry holes in the choice assays between corn and sorghum, and between sugarcane and corn. While corn has been known as a host of the Mexican rice borer for at least 84 yr, its role in area-wide population dynamics and control efforts has likely been greatly underestimated.     

植物MYB转录因子的同义密码子使用偏好性分析

前人研究显示MYB（V-myb avian myeloblastosis viral oncogene homolog）转录因子对应答植物逆境胁迫等有重要作用,甘蔗MYB转录因子的研究报道并不太多。通过对与甘蔗亲缘关系近的禾本科作物玉米、高粱和水稻MYB转录因子基因家族的密码子使用偏好性进行比较分析。结果表明,3个物种的MYB转录因子的蛋白质序列上存在高度的保守性,仅有部分密码子使用不太一样,此外,分析还显示基因碱基组成有可能在一定程度上影响了密码子的偏好使用。     

Evaluation of performance of sorghum varieties grown in Tokyo for sugar accumulation and its correlation with vacuolar invertase genes SbInv1 and SbInv2

ABSTRACT

The sugar-accumulating potential of global and local sweet and grain sorghum varieties were tested under the local conditions. The basis for this study was the dependency of sugar accumulation on temperature and photoperiod. Thus, the efficacy of cultivars as a bioenergy source would need to be determined based on their performance under the local environmental conditions. A strong correlation of sucrose content with brix was observed, enabling large-scale screening of varieties for high sucrose content. The morphological characteristics inherent in sweet sorghum, such as tall stems, greater number of leaves and a longer vegetative period, were found to correlate with the total stem sugar content. Assessment of sugars along the stem revealed maximum sugar accumulation in the upper intermediate to upper internodes in most of the varieties tested. The maximum theoretical ethanol yield (MTEY), a function of brix and juice yield, was determined as a better indicator of testing the performance of a variety as a potential source of bioethanol, mainly due to a negative correlation of stem juiciness and sucrose content in the varieties tested. Further, the relative expression of vacuolar invertase genes, SbINV1 and SbINV2, was studied, and a strong negative correlation of SbINV2 to stem sucrose content was observed. This reveals a possibility of involvement of vacuolar invertase gene, SbINV2, in sugar accumulation in sweet sorghum stems, and as a key candidate for molecular breeding studies for higher stem sugar content.     

国内外高粱深加工研究现状与发展前景

高粱既可直接食用、饲用，又可加工利用。近年来，国内外高粱深加工研究发展很快，除传统的用高粱作主食、酿制白酒、啤酒、做饲料等外，还对高粱的多种加工用途进行了有益的探索。如：用甜高粱茎秆造酒，用饲草高粱作青贮和青饲料，用高粱生产酒精，用高粱壳提取色素，用高粱籽粒提取淀粉，生产高粱面包、高粱甜点等等。高粱深加工发展前景看好。     

Application of very high gravity technology to the cofermentation of sweet stem sorghum juice and sorghum grain

Ethanol production from mixtures of sweet stem sorghum juice and sorghum grain was investigated under normal and very high gravity (VHG) fermentation conditions. Fermentation was carried out using Saccharomyces cerevisiae yeast strain N96 at 30°C. For VHG fermentation, sucrose was added to the sweet sorghum juice to obtain a concentration of 34 g per 100 ml of dissolved solids. Fermentation was carried out for 96 h using malted and unmalted milled sorghum grain from sorghum cultivars DC-75 and SV-2. Under VHG conditions, maximum ethanol levels were about 16.8% (v/v) and 11% (v/v) for media containing malted and unmalted milled sorghum grain, respectively. Although fermentation did not occur to completion, levels of ethanol obtained under VHG conditions were three times higher than the levels obtained under normal fermentation conditions. Under VHG conditions, about 8 g/100 ml of dissolved solids remained in the fermentation media after ethanol production had ceased while under normal fermentation conditions, about 4 g/100 ml of dissolved solids remained unused in the fermentation media. There was an initial decline in free amino nitrogen (FAN) levels up to 34 h followed by an increase up to 96 h under VHG fermentation conditions. Levels of assayable proanthocyanidins (PAs) from sorghum cultivar DC-75 were reduced during fermentation.     

Transgenic Bt corn varietal resistance against the Mexican rice borer,Eoreuma loftini (Dyar) (Lepidoptera: Crambidae) and implications to sugarcane

The Mexican rice borer, Eoreuma loftini (Dyar), attacks crops including corn, Zea mays L., rice, Oryza sativa L., sorghum, Sorghum bicolor (L.) Moench, and sugarcane, Saccharum spp. Strongly resistant varieties of any kind, native or otherwise, have not been identified. A field plot corn variety test using two transgenic Bt varieties, Pioneer 31G71, expressing the Cry1F insecticidal protein, and Golden Acres 28V81, expressing the Cry1A.105, Cry2Ab2, and Cry3Bb1 insecticidal proteins, and two non-Bt controls, Dekalb DKC 69-72 and BH Genetics 9050, all four commonly grown in the Lower Rio Grande Valley of Texas, showed that, although oviposition preference was not affected, 28V81 resisted larval stalk boring to the extent that Mexican rice borer injury was almost non-existent. Pioneer 31G71 was infested nearly as much as the controls, but larval development to adulthood was reduced by ≈70%. Rearing larvae on 5, 50, 500, and 5000 μg of corn leaf tissue per ml of artificial diet showed that, while the three lowest concentrations did not affect larval growth and development, the high concentration of 28V81 reduced survivorship to the pupal stage, decreased weight of 4-wk-old larvae, and prolonged development to pupation. Lower numbers of pheromone trap-captured adults at the edges of commercial Bt and non-Bt corn fields showed that populations were lower at the Bt cornfields, suggesting a lesser rate of adult production. Because corn is a preferred host plant over sugarcane, sorghum or rice, use of resistant transgenic Bt corn varieties will likely protect the crop from the substantial injury that can be caused by the pest. This study also suggests that Bt genes might result in similarly strong resistance when inserted in other vulnerable crops such as sugarcane.     

新型绿色可再生能源作物--甜高粱及其育种策略

甜高粱是一种新型的可再生能源作物，用甜高粱生产酒精已引起全世界的重视。在能源供应日益紧张的今天，不断研究和改革甜高粱育种方法、尽快将其应用到育种实践中去，选育出优良甜高粱杂交种，充分发挥其高能作物的优势，是极其必要的。发展甜高粱生产，将有效缓解能源危机，具有良好的经济、社会和生态效益。     

Impact of nitrogen fertilization on Mexican rice borer (Lepidoptera: Crambidae) injury and yield in bioenergy sorghum

The Mexican rice borer, Eoreuma loftini (Dyar) (Lepidoptera: Crambidae), is a serious pest of sugarcane (Saccharum spp.), sorghum (Sorghum bicolor (L.) Moench), corn (Zea mays L.), rice (Oryza sativa L.), and related graminaceous bioenergy crops. A two-year field study was conducted in Jefferson County, TX to examine the impact of nitrogen (N) fertilization on E. loftini infestations and subsequent yields in cultivars of high-biomass and sweet sorghum. In 2013, percentage of bored internodes and number of adult emergence holes per stalk increased with higher N rates; however, only the percentage of bored internodes was impacted by N in 2014. Yields from both years indicated that N rate was positively associated with increases in stalk weight and ethanol productivity, but not sucrose concentration. Because higher N rates were associated with increased yields despite having greater levels of E. loftini injury, our data suggest that increases in yield from additional N outweigh decreases from additional E. loftini injury. Fertilization rates maintained between the recommended 45 and 90 kg N/ha minimize risks of negative area-wide impacts from increased production of E. loftini adults, while still allowing for optimum yields.     

远缘杂交在水稻遗传育种中的应用

水稻种内基因资源的充分利用为我国粮食增产作出了巨大贡献。为进一步挖掘种间、属间及亚远缘物种的有利基因,维持栽培稻遗传多样性,实现水稻超高产、优质和多抗育种,本文综述了远缘杂交所涉及的分子机理及水稻远缘杂交育种的思路,概述了水稻种间和属间远缘基因资源的研究成果、远缘杂交障碍及克服途径、多种技术在水稻远缘杂交方面的发展概况等,为加快远缘物种基因资源在水稻育种上的应用提供参考。     

A simple method for squeezing juice from rice stems and its use in the high-throughput analysis of sugar content in rice stems

Sugar content in rice (Oryza sativa L.) stem is an agronomically important trait for rice used in straw silage or whole-crop silage. However, the mechanisms underlying sugar accumulation in rice stems remain unclear, mainly due to the time-consuming method for measuring sugar content. Here, we established a simple method for squeezing stem juice from rice plants, similar to that used during breeding selection in sugarcane or sorghum. The Brix value of the stem juice, which can easily be measured using a portable refractometer, significantly correlated with the soluble sugar contents in the stem juice and tended to correlate with those in stem tissues. This indicates that the Brix value of the stem juice can be used for estimating the sugar content in rice stems. This simple estimation method will be a useful tool for high-throughput analysis of sugar content in rice stems during mutant screening, QTL analysis, and breeding selection.     

甜高粱主要农艺性状遗传参数分析

对64份国内外甜高粱种质资源的9个农艺性状进行了遗传多样性等遗传参数研究。结果表明:各农艺性状多样性指数均较大,平均值为1.86,榨汁率多样性指数最小,为1.58,全生育期多样性指数最大,为1.99,表明64份甜高粱种质具有丰富的遗传多样性;穗长、全生育期、株高的遗传力较高,受环境影响小,可以在早世代选择;茎秆鲜重和穗长的遗传进度较高,获得的遗传增量大,直接选择效果较为理想;相关分析表明:除了穗长与茎秆鲜重相关不显著外,其它性状均与茎秆鲜重呈极显著正相关。株高与其它性状均呈极显著正相关。     

12个甜高粱品种在重庆地区的生产力和总糖含量表现

通过对12个供试甜高粱品种（组合）茎秆的鲜产量、总糖含量、总糖产量和籽粒产量的比较,结果为：茎秆鲜产量以A4为最高,达到41 385.0 kg/hm2,茎秆总糖含量以A9为最高,达到11.26%,茎秆总糖产量以A9为最高,达到4 285.4 kg/hm2,籽粒产量以A12为最高,达到2 723.4 kg/hm2。综合评价初步认为,在12个供试品种（组合）中,以A9组合和A3品种比较适合在重庆做生物质能源原料。     

Sorghum extrusion process combined with biofortified sweet potato contributed for high iron bioavailability in Wistar rats

This study aimed to evaluate the effect of sorghum and sweet potato on the bioavailability of iron, gene expression of proteins involved in iron metabolism and the plasma antioxidant capacity in animals fed with whole sorghum grains processed by dry heat or extrusion, combined or not with sweet potato flour with high content of carotenoids. Five experimental groups were tested (n = 7): dry heat sorghum flour (DS); extruded sorghum flour (ES); whole sorghum flour + sweet potato flour (DS + SP); extruded sorghum flour + sweet potato flour (ES + SP) and positive control (FS). The evaluations included: hemoglobin gain, hemoglobin regeneration efficiency, gene expression of divalente metal transporter 1 (DMT-1), duodenal citochroma B (DcytB), ferroportin, hephaestin, transferrin and ferritin and total plasma antioxidant capacity (TAC). The ES + SP group showed higher (p < 0.05) expression of DcytB, ferroportin and hephaestin when compared to the control group. The DS group showed high (p < 0.05) expression of DMT-1 and the ES showed high mRNA expression of transferrin and ferritin. The changes in the sorghum physicochemical properties from extrusion process reduced the iron and phytate content, and increased the gene expression of proteins involved in iron metabolism, improving iron bioavailability. The combination of sweet potato and sorghum flour (dry or extruded) improved the iron capture and total antioxidant capacity, probably due to the presence of β-carotene and antioxidant compounds.     

能源型甜高粱分期播种试验初报

以甜高粱品种辽甜1号和辽甜3号为试验品种，在南宁地区进行种植，对其不同播期的农艺性状、产量性状及茎秆汁液品质进行调查分析。结果表明：4月底播期，辽甜1号、辽甜3号的茎秆产量和汁液锤度在各播期里最高，分别达4 329.11 kg/亩（1亩=666.7 m2，下同）、5 340.28 kg/亩和21.18％、18.27％，是这2个品种在南宁地区的最佳种植时期。3月到8月播期，茎秆产量、籽粒产量及汁液锤度都表现良好，是这2个品种在南宁地区的适宜种植时期。     

Biomass, extracted liquid yields, sugar content or seed yields of biofuel feedstocks as affected by fertilizer

Harvesting products from plants for conversion into renewable resources is increasing in importance. Determination of nutrition requirements for the applicable crops is necessary, especially in regions where the biofuel feedstock crops have not been historically grown. Sunflower (Helianthus annuus L.), two hybrids and one variety; sweet and grain (milo) sorghums (both Sorghum bicolor L.), one variety each, and sweet corn (Zea mays var. rugosa Bonaf.), four cultivars, were provided the recommended and twice the recommended rate of fertilizer. Biomass, expressed liquid volumes and sugar contents of sweet sorghum and sweet corn were determined. Grain yields of milo and sunflower and oil content of sunflower were determined. Sweet corn stalk sugar levels were below what is expected from field corn (maize), and were not affected by fertilizer rate. Sweet sorghum biomass and sugar content were within expected ranges and not affected by fertilizer rate. Milo grain yields were higher with increased fertilizer. Seed yield in Sunflower, which was below expected levels, was inconsistently affected by fertilizer rate, years or varieties. Overall crops year and cultivar/variety had more effect on results than did fertilizer. There does not appear to be a reason to provide fertilizer above recommended rates in production of these crops.     

3种生长延缓剂对甜高粱幼苗生长和生理特性的影响

采用不同浓度的多效唑、矮壮素、比久对甜高粱种子进行浸种处理,结果表明,3种生长延缓剂中,多效唑对甜高粱幼苗生长的抑制作用最强;0.1～0.2 g/L的多效唑浸种处理可以提高种子的发芽势和发芽率,但幼苗生长受到过度抑制而表现不正常;5g/L和10g/L的矮壮素和比九处理可以延缓甜高粱幼苗生长,使叶片缩短、厚度增加,具有提高幼苗可溶性糖、可溶性蛋白质和叶绿素含量的作用,促进了幼苗对干物质的合成和积累,使幼苗十物质重显著提高,有利于培育壮苗.其中,10g/L矮壮素浸种处理效果最佳.     

生长素调控因子OsGRF4协同调控水稻粒形和稻瘟病抗性

【目的】水稻粒形为多基因控制的复杂数量性状,同时影响稻谷产量和稻米外观及碾磨品质,挖掘粒形相关基因并解析其遗传机制,对于水稻高产和优质品种选育具有重要意义。【方法】以前期利用大粒籼稻品种特大籼(TDX)为供体亲本、小粒粳稻品种日本晴(Nipponbare,NPB)为轮回亲本获得的一个大粒近等基因系,在水稻第2染色体上初定位粒形调控基因GS2.2(grain size 2.2)的基础上,对GS2.2基因进行了精细定位和候选功能基因的克隆鉴定。【结果】利用BC₄F₂群体中2887份极小粒单株及该群体中70份基因型杂合的大粒单株自交获得的BC₄F₃群体,将GS2.2基因精细定位在2M-7-1与2M-9之间的160.6 kb的区间内。该区间编码18个基因开放阅读框(ORF1-18)。测序发现ORF18基因在大粒近等基因系与小粒日本晴等位基因间存在5个SNP差异,ORF18编码生长素调控因子蛋白OsGRF4。采用CRISPR/Cas9基因编辑技术对大粒近等基因系中ORF18进行敲除,发现ORF18敲除株系粒长变短,证实ORF18是GS2.2的功能基因。进一步对ORF18大粒近等基因系和基因编辑敲除株系进行稻瘟菌室内离体和病圃接种鉴定,发现ORF18大粒近等基因系稻瘟病抗性增强,而基因编辑敲除株系稻瘟病抗性减弱,表明ORF18正调控水稻稻瘟病抗性。【结论】本研究发现的生长素调控因子OsGRF4协同调控水稻粒形和稻瘟病抗性,为协调水稻高产和抗性育种提供了重要的基因资源。     

Genome-Wide Survey and Expression Analysis of P1B-ATPases in Rice,Maize and Sorghum

P_1B-type ATPase ion pumps that transport heavy metal ions across cellular membranes are essential for plant growth and development. To date, a genomic comparison overview of the family in rice, maize and sorghum is not yet available. In this study, a total of 31 heavy metal P_1B-type ATPase (HMA) genes were identified, including 9 in rice, 11 each from maize and sorghum. They were classified into two distinct subfamilies based on their sequence composition and phylogenetic relationship. Four pairs of HMA genes were expanded via gene duplication with tandemly duplicated. Comprehensive analyses were performed to investigate the expression profiles of HMA genes in various tissues by using quantitative real-time PCR. Some HMA members exhibited abundant and tissue-specific expression patterns. Moreover, most of the genes were found to be differentially expressed under the Cu/Cd treatment. This study will facilitate further studies on P_1B-type ATPase family and provide valuable hints for the functional validation in rice, maize and sorghum.