Genetic Control of Five Isozyme Systems in Sugar Beet (Beta vulgaris L.)

Genetic markers are most useful in plant breeding but so far only a few morphological and isozyme markers are known in Beta vulgaris. In this paper, the isozyme systems aconitase (AGO), NADH-:dihydrolipoamiddehydrogenase (DIA), esterase (EST), fructose-1,6-bisphosphatase (FDP), and hexokinase (HK) were genetically analysed. Isoelectric focussing (IEF), vertical polyacrylamide gel electrophoresis (PAGE) and starch gel electrophoresis (SGE) were used to separate the isozymes. A total of eight loci could be identified. The allelic forms of Aco-1, Dia-1, Est-1, Est-5 and Hk-1 are active monomers. The gene products of Est-3 and Fdp-2 are active as dimers. Est-2 also showed allelic forms without activity; null-alleles.     

栽培甜菜雌配子体发育中的超微结构

应用透射电镜技术研究栽培甜菜(Beta vulgaris L.)雌配子体发育过程的超微结构特征, 丰富被子植物生殖生物学方面的资料, 并为甜菜相关研究提供借鉴。观察结果表明,栽培甜菜雌配子体发育类型为蓼型。功能大孢子时期核糖体密集, 线粒体和质体分裂以增加数目;单核胚囊时期, 细胞体积增大, 小液泡融合为2个大液泡,分别位于珠孔端和合点端,细胞核增大, 核仁显著, 至发育后期核膜呈微裂齿状,细胞器数量不断增加,珠孔端形成明显的壁内突;二核胚囊时期, 胚囊迅速扩张, 形成中央大液泡, 胞质中细胞器增多;四核胚囊时期除细胞器的数量继续增加外,质体中开始出现淀粉粒;八核胚囊时期相当短暂, 很快细胞化,初期的七细胞八核胚囊中各细胞均以初生壁相隔, 壁上存在胞间连丝, 沿细胞壁分布着众多伸展状粗面内质网和活跃分泌小泡的高尔基体;细胞化后期, 卵、助细胞出现极性分化, 胞质中的众多小泡与细胞膜融合, 参与细胞壁及丝状器的建成;中央细胞在珠孔端与胚囊壁相邻的部位形成壁内突。栽培甜菜雌配子体发育进程中胚囊体积不断增大,细胞器种类与数量呈增加趋势,表明各时期代谢较活跃。在功能大孢子及单核胚囊早期是通过合点端的胞间连丝与珠心细胞以共质体形式相通;在单核胚囊发育后期至细胞化胚囊,是通过珠孔端所形成的壁内突来扩展质膜表面积, 以加强非共质体之间物质的运输。     

栽培甜菜雌配子体发育中的超微结构

应用透射电镜技术研究栽培甜菜(Beta vulgaris L.)雌配子体发育过程的超微结构特征, 丰富被子植物生殖生物学方面的资料, 并为甜菜相关研究提供借鉴。观察结果表明,栽培甜菜雌配子体发育类型为蓼型。功能大孢子时期核糖体密集, 线粒体和质体分裂以增加数目;单核胚囊时期, 细胞体积增大, 小液泡融合为2个大液泡,分别位于珠孔端和合点端,细胞核增大, 核仁显著, 至发育后期核膜呈微裂齿状,细胞器数量不断增加,珠孔端形成明显的壁内突;二核胚囊时期, 胚囊迅速扩张, 形成中央大液泡, 胞质中细胞器增多;四核胚囊时期除细胞器的数量继续增加外,质体中开始出现淀粉粒;八核胚囊时期相当短暂, 很快细胞化,初期的七细胞八核胚囊中各细胞均以初生壁相隔, 壁上存在胞间连丝, 沿细胞壁分布着众多伸展状粗面内质网和活跃分泌小泡的高尔基体;细胞化后期, 卵、助细胞出现极性分化, 胞质中的众多小泡与细胞膜融合, 参与细胞壁及丝状器的建成;中央细胞在珠孔端与胚囊壁相邻的部位形成壁内突。栽培甜菜雌配子体发育进程中胚囊体积不断增大,细胞器种类与数量呈增加趋势,表明各时期代谢较活跃。在功能大孢子及单核胚囊早期是通过合点端的胞间连丝与珠心细胞以共质体形式相通;在单核胚囊发育后期至细胞化胚囊,是通过珠孔端所形成的壁内突来扩展质膜表面积, 以加强非共质体之间物质的运输。     

Root Yield and Quality of Sugar Beet (Beta vulgaris L.) in Relation to Plant Population

Introduction of genetic monogerm seed and the advancement of agricultural techniques raised the possibility of eliminating costly thinning operations in sugar beet farming. However, this requires high and uniform seedling emergence to provide prerequisite plant populations in the field. This work investigated the minimum levels of seedling emergence for non-thinned sugar beet farming. The experiment was carried out in a loamy soil to investigate the effects of three levels of emergence (60, 50 and 35%) and two intra-row spacings (8 and 15cm) with or without thinning using seed source of graded viability of cv Eva in 1992 and 1993 under field conditions in Eastern Anatolia. Plant population throughout the growing period was monitored and data were taken of yield and quality parameters. On average, 60, 50 and 35% emergence levels were achieved using seed source of 90, 72 and 54% viability range, which resulted in different levels of plant stands depending on intra-row spacing and thinning. Plant density at harvest ranged between 49 500 and 124 500 ha⁻¹ on average depending on initial establishment, intra-row spacing and thinning. Root yield and sugar yield were the highest at plant stands between 90 000 and 100 000 ha⁻¹, below or above of which declined. Impurity contents (e.g. Na, K and a-amino N contents) and extractable sugar content decreased but sugar content and clear juice purity enhanced as plant stands at harvest increased.     

氮素形态对甜菜氮糖代谢关键酶活性及相关产物的影响

用营养液培养的方法, 以甜研7号为试验材料, 研究了同一氮素水平不同氮素形态比例对甜菜氮、糖代谢关键酶: 硝酸还原酶(NR)、谷氨酰胺合成酶(GS)和蔗糖合成酶(SS)活性及相关产物的影响. 结果表明叶片中NR活性随NO-3比例增加而增加; 根和叶片中GS活性, 当NH+4小于NO-3时, 随NH+4比例增大而增加, 当NO-3∶NH+4为1∶     

甜菜花叶病及种质资源抗性的研究进展

甜菜花叶病是甜菜生产的世界性病害,可经多种蚜虫非持久性传播。花叶病毒侵染甜菜,嫩叶常出现脉间褪绿的明脉现象,发生皱缩;功能叶片会出现明暗相间的绿色斑驳,进而产生花叶,严重的发生坏死,给甜菜生产造成巨大损失。归纳了BtMV病原及种质资源抗性的研究概况,分析了BtMV病原生物学特性、病原基因组及编码蛋白结构功能,病原介体及病毒复制模式,以及BtMV侵染甜菜植株及叶片的结构与微观变化。总结了当前用于甜菜花叶病的检测方法和甜菜种质资源的花叶病抗性与抗病反应的研究进展。提出了当前甜菜抗花叶病需要深入开展的研究论题,并对甜菜抗花叶病分子机理和基因工程育种进行了展望。为该病的深入性研究,特别是基因工程育种提供参考。     

不同温度对甜菜种子吸胀过程中线粒体CCO和MDH活性的影响

吸胀温度显著地影响甜菜种子的萌发速率。吸胀温度为３０℃和２０℃时,５０％萌发所需要的时间分别为３６ｈ和５６ｈ;在７℃中吸胀１２０ｈ,仍未见萌发。在种子吸胀过程中,粗线粒体蛋白的含量,细胞色素Ｃ氧化酶和苹果酸脱氢酶的活性增加,且它们在３０℃和２０℃中的活性较高,在７℃下较低。     

Seasonal Development of Genotypic Differences in Sugar Beet (Beta vulgaris L.) and Their Interaction with Water Supply

Depending on genotype, sugar beet can differ considerably in yield and quality characteristics. These are additionally modified by environmental conditions with drought stress recently gaining in importance, restricting growth and altering the chemical composition of the beet. The occurrence and development of these genotypic differences during the vegetation period and their possible interaction with environmental conditions were investigated. In 2002 and 2003, four sugar beet genotypes differing in yield and quality and putative different with regard to drought tolerance were tested in field trials, partly under irrigated conditions, in a total of 10 environments with consecutive harvests starting in early summer. In 2 years of stress and non-stress conditions they exhibited significant differences for taproot and leaf dry matter and the concentration of sucrose, K, Na and α-amino nitrogen in the taproot. These differences existed already in mid-June and virtually did not change any more from this time on. Accordingly, interactions between genotype and harvest date did not occur. For sugar beet, genotype by environment interactions generally do not exist. Water supply, as an important single determinant of the effect of the environment, was studied separately analysing data from selected locations. Under drought conditions, withholding irrigation reduced leaf and taproot growth and root-to-leaf ratio, decreased the percentage of sucrose in dry matter and resulted in an accumulation of α-amino N. Interactions between genotype and water supply did not occur for any of the parameters under study. A genotype-specific high α-amino N content, which might be of advantage for osmoregulation, did not improve the adaptation to drought. Differences in leaf maintenance or taproot-to-leaf ratio during drought also did not affect yield response. Due to the lack of interaction between genotype and harvest date as well as between genotype and irrigation it is concluded that harvest date or climatic factors of the growing region do not have to be taken into consideration when choosing a variety.     

A DNA Probe for Rapid Screening of Sugar Beet (Beta vulgaris L.) Carrying Extra Chromosomes from Wild Beets of the Procumbentes Section

Genes from wild species of the Procumbentes section can be transferred to sugar beet chromosomes via translocations. Since large translocations, including for example a gene for nematode resistance, generally result in meiotic disturbances, there is a need to select new diploid resistant beets from progenies of monosomic wild beet addition lines. A dispersed repetitive DNA probe, which is closely correlated with the resistance gene and hybridizes exclusively with wild beet DNA, appears to be highly superior to selection based on isozyme markers. Characteristic ‘fingerprints’ on the available monosomic addition lines reveal the existence of at last 5 different chromosomes in the wild species each housing a gene for nematode resistance. This probe can be used advantageously to identify individuals carrying the intact or fragmented wild beet chromosomes, or even various amounts of translocated chromatin. Strategies are discussed for the identification of new translocation types using straightforward squash dot or Southern hybridization techniques in combination with the wild beet DNA probe.     

普通小麦及近缘粗山羊草α-醇溶蛋白基因的克降、定位与进化分析

通过特异PCR引物设计,从普通小麦品种(豫麦34和烟农19)和粗山羊草(T9、T197、T48、T176和T17)中扩增、克隆了7个新的α-醇溶蛋白基因,分别命名为Gli-YM34、Gli-YN19、Gli-T9、Gli-T197、Gli-T48、Gli-T176和Gli-T17,基因序列长度为846~891 bp,编码282~297个氨基酸残基,都具有α-醇溶蛋白的典型结构特点。其中Gli-YM34和Gli-YN19基因推导的醇溶蛋白都含有一个额外的半胱氨酸残基,可能对面筋品质有正向作用。根据α-醇溶蛋白氨基酸序列所具有的4种T细胞抗原表位和多聚谷氨酰胺重复区的平均长度以及中国春缺体四体分析,将来自普通小麦品种的Gli-YM34和Gli-YN19基因定位在6D染色体上的Gli-D2位点,而且Gli-YM34和Gli-YN19与来自粗山羊草的α-醇溶蛋白基因具有很高的序列相似性,进一步证明粗山羊草是普通小麦D基因组的供体。在克隆的4个典型α-醇溶蛋白基因中检测到21个SNP和1个9 bp的缺失。系统进化分析表明,α-醇溶蛋白基因与低分子量谷蛋白亚基基因关系较近,在大约43.69百万年时分化,与ω-醇溶蛋白和HMW-GS基因亲缘关系较远,它们的分化时间大约为79.39百万年。     

甜菜nia基因的克隆及不同氮素形态诱导的差异表达

利用同源序列克隆方法从二倍体甜菜品种Ty7中获得氮素诱导nia基因片段,通过RACE技术克隆nia基因全长序列,该基因ORF长度2 718 bp,编码905个氨基酸,并已在GenBank上登录(EU163265),基因组中nia以低拷贝数存在。nia编码蛋白的等电点为6.12,推测分子量为102 kD,以NADH为电子供体。为揭示不同氮素形态和处理对甜菜nia基因表达的影响,采用半定量PCR方法检测不同氮素形态诱导nia基因mRNA的表达,同时测定酶活力。结果表明,当铵态氮诱导nia基因时,低浓度的铵离子能促进基因的表达,过高浓度的铵离子抑制基因的表达。当硝态氮诱导nia基因时,随处理浓度的增加,nia的表达加强,呈正相关关系。用30 mmol L^–1硝态氮诱导4 h后,nia基因表达达最高值,约在6 h后,表达明显下降。     

Leaf Carbon Isotope Discrimination and its Relation with Qualitative Root Traits and Harvest Index in Sugar Beet (Beta vulgaris L.)

Twenty commercial sugar beet varieties were grown at two sites in central Greece during the 2001 growing season (March to October), in a completely randomized block design with three replications. Carbon isotope discrimination (Δ) in sugar beet leaves was found to be affected by site and marginally by variety. No relation was found between fresh root yield and Δ, but significant correlations were found between Δ and sucrose content (positive) and between Δ and root α‐amino‐N (negative). Also, the harvest index, determined at one site, was negatively correlated with Δ.     

Random Amplified Polymorphic DNA (RAPD) Markers in Sugar Beet (Beta vulgaris L.): Mapping the Genes for Nematode Resistance and Hypocotyl Colour

The random amplified polymorphic DNA (RAPD) technique was adapted for segregation analysis in sugar beet. 83 I_O/I individuals were scored with a set of 20 arbitrary decamer primers. 4 preliminary linkage groups could be established, enclosing 9 RAPD markers, 2 isozyme loci, a gene for the hypocotyl colour and a gene for resistance to the root knot nematode (Heterodera schachtii Schm.).     

Correlation between polyovules and number of pistil lobes (stigmatic rays) in sugar beet (Beta vulgaris L.)

The polyovules phenomenon in sugar beet has the same importance for breeding as multigermity, because only mono-ovulic monogerm plants can be used for mechanized cultivation. The polyovules trait was investigated in two Japanese lines, mono-ovulic line NK-219-O and polyovulic line NK-183-cms. Plants of the former had three pistil lobes (stigmatic rays) in each flower, a single ovule per ovary, a single seed in the mature fruit and a single seedling germinated in subsequent tests. Plants of the latter had 5–6 pistil lobes in each flower, 2–3 ovules per ovary, double-seeds and double-seedlings. The correlations between a Pistil Lobes Index (PLI) and thenumber of ovules per ovary, percentage of double-seeds inan X-ray test, and percentage of double-seedlings in germination tests were 0.885, 0.838, and 0.749, respectivelyindicating close associations. These results are important both in sugar beet breeding for indirect selection to eliminate the undesirable polyovules trait and for botanicalinvestigations of the relationship among different parts of the gynoecium.     

栽培甜菜中央细胞受精前后的超微结构

应用透射电镜技术比较栽培甜菜(Beta vulgaris)中央细胞受精前后的超微结构特征,以完善甜菜生殖生物学研究,并为相关研究提供基本资料。结果表明,中央细胞在受精前后的超微结构变化主要表现于核及核周围胞质。中央细胞的2个极核融合较早,在花蕾时期即以次生核形式存在,具双相核仁,可明显分辨纤维区和颗粒区;有的有额外小核仁和核仁液泡。核周围的细胞质中具丰富的细胞器且功能活跃,包括线粒体、质体(含或不含淀粉粒)、高尔基体、核糖体和粗面内质网。受精后,形成初生胚乳核,核周围胞质中变化最大的是质体,形成众多变形质体,形态多样,且富含淀粉粒。初生胚乳核分裂过程中,核仁消失,核膜崩解为囊泡状结构,粗面内质网减少,滑面内质网增加。分裂形成2个胚乳游离核,核周围胞质与初生胚乳核相似。总之,中央细胞在受精前后的超微结构特征均呈现代谢活跃状态。     

An Extended Linkage Map of Sugar Beet (Beta vulgaris L.) Including Nine Putative Lethal Genes and the Restorer Gene X

An extended genetic map of sugar beet (Beta vulgaris L.) is presented encompassing 177 segregating markers (2 morphological traits, 7 isozymes, and 168 RFLP markers) on 9 linkage groups. The linkage map comprises 1057.3 cM equivalent to an average genetic spacing of 6.0 cM/marker. The length of individual linkage groups varies between 80.7 (group VIII) and 167.4 cM (group VIII). The number of markers per linkage group ranges between 13 and 24. No indication of duplicate regions was found, confirming the true diploid nature of B. vulgaris. Twenty-six markers (15 %) deviated significantly (a = 0.01) from the expected segregation ratio. This distorted segregation was probably caused by linkage with lethal genes. Four such genes (designated Let Ib, Let 5b, Let 6b, Let 8) could be located at discrete positions due to their absolute linkage to skewed RFLP markers. The restorer gene X has been located terminally on linkage group ÜI, 9.6 cM distant from RFLP marker pKP1238.     

内生真菌对甜菜主要农艺性状及氮糖代谢关键酶活性的影响

采用内生真菌F11液浸种、喷叶及灌根处理方法,调查其对甜菜栽培品种KWS2409的主要农艺性状及对甜菜氮、糖代谢关键酶即硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、蔗糖合酶(SS)和蔗糖磷酸合酶(SPS)活性的影响。结果表明,内生真菌F11菌株对甜菜的含糖量有明显的提高作用,其中以灌根处理效果最好,其叶鲜重、叶绿素含量、单根重、含糖率和产糖量的平均值分别提高了66.67%、47.42%、6.96%、17.46%和25.63%。在整个生育期,内生真菌F11显著提高了氮糖代谢酶活性,其中NR和GS活力分别呈“M”型双峰曲线和抛物线型变化,而SS和GS活力呈单峰曲线变化,后期根部SS合成活力明显高于分解方向活力,生育前期SPS活力高于后期。叶丛形成期达到最高峰,说明NR、GS、SS和SPS活性的增强是甜菜含糖量升高的主要生理原因。     

Variation in some agronomically important characters in a germplasm collection of beet (Beta vulgaris L.)

Summary A geographically representative selection of germplasm of Beta vulgaris, section Beta has been assessed for characteristics important in sugarbeet breeding, including downy mildew resistance, resistance to aphid colonisation and infection by the beet virus yellow complex. The occurrence of maintainer lines for cytoplasmic male-steriles was also investigated. Desirable qualities were found in some accessions, including nothern European wild vulgaris ssp. maritima and some old multigerm cultivars of fodder beets.     

甜菜蔗糖代谢两种相关酶的活性变化及其相互关系

为了研究不同类型甜菜品种蔗糖代谢能力存在明显差异的生理机制,以糖用甜菜(高糖)和饲用甜菜(丰产)为试验对象,测定了不同生育阶段叶片、叶柄和块根中的蔗糖合成酶(SS)和蔗糖磷酸合成酶(SPS)活性。结果表明:叶片、叶柄和块根中存在蔗糖合成与降解过程,不同类型甜菜在生长期蔗糖合成酶和蔗糖磷酸合成酶活性存在差别,同一品种不同部位,块根中蔗糖的代谢强度高于叶片和叶柄,糖用甜菜两种酶活性高于饲用甜菜,蔗糖代谢相关酶活性差异是品种特性,可用于甜菜的选种实践。