Inheritance of 'Hullability' in Sunflowers (Helianthus annuus L.)

Improvement of the feed value of sunflower seedmeal by hulling achenes before crushing is of economic importance and varieties with good ‘hullability’ (ease of hulling) would be economically significant. Knowledge of the genetics of this characteristic is therefore necessary. Studies of heritability were made on a factorial cross of six female and six restorer lines. Hullability was measured as the ratio of hull removed by a laboratory huller against the total quantity of hull in the achene. The genotypes studied showed a wide range of hullabilities. There were considerable environmental effects but only small interactions; correlations between results in different locations were significant. Additive genetic effects predominated, although there were some interactions between parental lines. Narrow-sense heritabilities were 0.73–0.85 for genotype means, 0.52 when calculated from individual plants, and 0.36–0.76 from parent-progeny regressions. Early selection based on progeny means or a combination of individual and progeny values would appear to be the best method of breeding for improved hullability of sunflower achenes.     

蜂蜜菊芋酒发酵工艺的研究

以菊芋为主要原料,蜂蜜补充糖源,对蜂蜜菊芋酒发酵条件进行研究。结果表明,三种不同加糖方式中,采用初始糖度20%(还原糖含量184.2 g·L~(-1)),糖度降为10%(还原糖含量92.90 g·L~(-1))后蜂蜜补糖至15%(还原糖含量146.3 g·L~(-1))的加糖方式进行酒精发酵,可明显提高酒体酒精度,降低残糖量;主发酵结束后酒精度为14.4%(V/V),还原糖含量降至8.00 g·L~(-1)。筛选出蜂蜜菊芋酒的发酵条件为:酵母菌接种量0.8 g·L~(-1),发酵温度26℃,初始p H 4,装液量70%,在此发酵条件下,酒体酒精度为14.0%(V/V),还原糖含量10.4 g·L~(-1),总酸含量6.90 g·L~(-1),按此工艺得到蜂蜜菊芋酒色泽金黄,香气浓郁,口感柔和。     

Interspecific hybridization between sunflower and wild perennial Helianthus species via embryo rescue

Interspecific crosses were made between the cultivated sunflower inbred line HA89 and accessions of five wild perennial Helianthus species (H. giganteus L., H. laevigatus T. & G., H. resinosus Small, H. pauciflorus Nutt. and H. decapetalus L.) resistant to broomrape ( Orobanche cernua Loefl.). Using the genetic male-sterile isogenic version of that line as female, successful reciprocal crosses were also obtained with all these wild species except with H. decapetalus. Five-day-old hybrid embryos were excised and cultured in vitro. In all cases, few mature plants were obtained from embryos in early developmental stages (early heart and globular) but up to 28% mature plants were obtained from embryos in later stages of development. Very immature embryos were difficult to excise without damage. Hybrid embryos and mature plants were obtained from all five wild species. Interspecific hybrid embryos from different wild species showed distinct developmental potentials, the proportion of hybrid embryos in different developmental stages varying among species. Differences in the proportion of hybrid embryos at the several developmental stages were also obtained for the reciprocal crosses (cultivated × wild), which showed higher proportion of fully developed embryos. Hybrids involving H. giganteus and cultivated sunflower were difficult to obtain without the use of embryo culture. Embryo culture proved to be an useful tool to overcome post-zygotic hybrid incompatibility in different Helianthus spp., and facilitated interspecific transfer of resistance to O. cernua. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of different microsatellite motifs for analysing genetic relationships in cultivated sunflower (Helianthus annuus L.)

A total of 16 different simple sequence repeat motifs and an M13 repeat sequence were used as hybridization probes in order to examine the molecular relationships between two German inbred and eight North American sunflower lines of known pedigree, and to determine the suitability of the individual short tandem repeats for genetic analyses and their occurrence in the sunflower genome. Only the oligonucleotides (ACA)₆, (CAT)₆, (CATA)₅, (GACA)₄ and (GATA)₄ proved to be well suited for the generation of scorable fingerprinting patterns, while the other (microsatellite) sequences were either too abundant or too rare. Although different levels of polymorphism were present in hybridizations with the different simple sequence repeats (SSRs), a clear separation of the German material from the USDA Hnes was feasible not only by evaluating 20 probe-enzyme combinations, but also by combining every individual probe with four different restriction enzymes. In the American material, the expected relationships could be proved with the single exception of a line that did not group in accordance with its pedigree.     

向日葵S-腺苷甲硫氨酸合成酶基因克隆与分析

为了研究S-腺苷甲硫氨酸合成酶(S-adenosylmethionine synthetase,SAMS)在向日葵(He-lianthus annuus)抗旱和耐盐过程中的作用,先根据计算机辅助克隆结果设计引物,抽提盐胁迫向日葵叶片的总RNA,然后采用RT-PCR扩增技术克隆了向日葵的1个SAMS基因(命名为HaSAMS1),HaSAMS1基因的编码序列长1173bp,编码390个氨基酸残基。HaSAMS1没有跨膜结构域,没有信号肽,含有SAMS蛋白的特征序列。HaSAMS1与其他物种的SAMS具有较高的序列相似性,与茼蒿(Chrysanthemum coronarium)和拟南芥(Arabidopsis thaliana)等高等植物SAMS的氨基酸序列一致性介于86.8%~96.9%。HaSAMS1基因的克隆为进一步研究向日葵抗旱和耐盐机理奠定了基础。     

Effect of wild Helianthus cytoplasms on agronomic and oil characteristics of cultivated sunflower (Helianthus annuus L.)

Sunflower (Helianthus annuus L.) productions reliance on a single source of cytoplasmic male sterility, PET1, derived from H. petiolaris Nutt., makes the crop genetically vulnerable. Twenty diverse cytoplasmic substitution lines from annual and perennial wild species were compared with the inbred line HA89 over four environments. Pairwise comparisons of alloplasmic male fertile (MF) with euplasmic MF lines suggested that cytoplasm of perennial H. angustifolius increased lodging while perennial cytoplasms of H. mollis, H. grosseserratus, and H. divaricatus reduced head size and consequently reduced yield and will need to be used with caution. Lines having annual species, cytoplasms had no effects on agronomic traits. Comparisons of CMS HA 89 with 10 CMS lines suggested yield‐reducing cytoplasmic effects of perennial H. maximiliani and annual H. annuus PI 413178 and PI 413024. Cytoplasmic effects on oil percentage and fatty acid composition were minimal and should not be of concern for sunflower breeders. In general, most cytoplasms of wild annual Helianthus species accommodate cultivated nuclear genes without drastic adverse interactions, and are potential sources of cytoplasmic diversity for sunflower breeding.     

Doubled haploid production of sunflower (Helianthus annuus L.) through irradiated pollen-induced parthenogenesis

Induced parthenogenesis as a possible method of haploidization in sunflower (H. annuus L.) was tested in previous investigations (Todorova et al., 1994) and conditions for reproducible regeneration of gynogenic doubled haploids were established by the present work. Forty eight treatments were studied, involving four pollen donors and four recipient hybrids. Pollen was irradiated with doses of 300 Gy, 600 Gy and 900 Gy. In total, 2279 embryos were cultivated in vitro of which 1107 plants were obtained and 582 of them produced seeds after selfing. The ploidy level of the regenerants was evaluated at the two – three leaf stage and 296 of the plantlets obtained were haploids. Some of them underwent spontaneous diploidization, the others were treated with colchicine solution for chromosome doubling. The diploid plantlets were checked for their gynogenic origin by genetical and biochemical methods. The effectiveness of the method expressed as the number of agronomically useful DH lines to the number obtained that were fertile and resistant to downy mildew, branched and unbranched plants was 8,6% on average. This revised version was published online in July 2006 with corrections to the Cover Date.     

不同品种油葵水浸提液对3种药用甘草化感效应的研究

为探讨油葵对甘草潜在的化感作用,选择矮大头(567 DW)、矮早丰(BKZ 9806)、新葵4号以及新葵6号等不同品种油葵和3种药用甘草为研究对象.用油葵根、茎、叶等器官不同浓度的水浸提液分别处理乌拉尔甘草、胀果甘草和光果甘草的种子和幼苗,比较0,25,50 mg/mL 3个浓度的水浸提液对3种甘草种子萌发和幼苗生长影响格局的差异.结果表明,不同油葵品种、不同油葵器官的水浸提液的化感效应不尽相同,矮大头浸提液对胀果甘草的种子萌发和幼苗生长均无明显抑制作用,并且其根的提取液显著促进了根的生长,在25,50 mg/mL浓度下分剐比ck高出26％和37％(p＜0.05),RI分别为0.21和0.27.根据本实验结果,在生产实践中建议选用矮大头与胀果甘草进行套种,可望能提高种植收益.     

Genetic variability in plants regenerated from in vitro culture of sunflower (Helianthus annum L.)

In vitro regenerated sunflower (Helianthus annuus L.) plants (R₁) were self-fertilized and the R₂ generation was evaluated for qualitative traits. A broad range of phenotypic variation was observed and mutation frequencies were calculated. Some in vitro induced variant phenotypes were similar to known spontaneous or induced mutations in sunflower, while others were new. Chlorophyll and carotenoid deficiencies, chimaerical variegation, fasciated stem and capitulum, abnormal shoot development, and other morphological variations, were noted. Substitution of anthers with petaloid structures in a disk-floret mutant indicates a possible homeotic mutation induced by in vitro tissue culture.     

Characterization of a green non‐dormant sunflower (Helianthus annuus) mutant NDG

A sunflower (Helianthus annuus L.) mutant was observed in the progeny of a cross between the sunflower cultivar ‘HA 89’ and an amphiploid of a Helianthus divaricatus L. × P21 cross that exhibited loss of dormancy induction in the developing embryo. Seeds of this mutant frequently germinate on the head about 40 days after pollination. The cotyledons of this mutant remain green, whereas some other non‐dormant mutants exhibit loss of pigmentation. The objectives of this investigation were to compare levels and activities of abscisic acid (ABA), a plant hormone that induces dormancy in developing embryos, in the non‐dormant green mutant (NDG) and ‘HA 89’ from which NDG was derived. Immunoassays showed that abscisic acid was present in NDG and the levels were not significantly different from those in ‘HA 89’. Exposure of excised NDG mutant embryos to 40 μm abscisic acid failed to prevent germination, suggesting that non‐dormancy could result from impairment in ABA receptors or from a defect in other proteins participating in the subsequent signalling pathway that normally induces dormancy.     

基于DUS测试性状的向日葵品种遗传多样性分析

利用NY/T 2433-2013中的43个基本测试性状对巴彦淖尔分中心的63个向日葵参试品种进行遗传多样性分析,并与32个近似品种作比较,结果表明：参试品种表型性状中共检测到124个等位变异,平均每个性状检测到3.0244个,变幅为1~8个。Shannon’s多样性指数（H＇）平均值为0.5582,变幅为0~1.8101,41个有效测试性状中有17.07%的性状存在5个及以上等位变异,有51.22%的性状存在3个及以上等位变异,参试品种的等位变异数、每个性状的等位变异数及其变异幅度、多样性指数平均值等均大于近似品种。在外观形态上,参试品种的遗传多样性比近似品种更丰富。UPGMA聚类分析发现,在相似系数为0.860时,可将95个向日葵品种分为2个类群。主坐标分析结果与UPGMA聚类分析结果基本一致,但是可以通过向日葵品种在主坐标中的位置更加直观地判断95个向日葵品种间的遗传相似性。总体看来,参试品种与近似品种形态差异较小,建议育种家拓宽向日葵亲本选育材料,从而促进向日葵材料创新及新品种选育。     

油葵品种在盐碱地的生态适应性研究

油葵品种在盐碱地的生态适应性研究结果表明,8个油葵品种生育期为113～116d,相差不大,在当地条件下均能正常成熟;各品种以A15产量最高,达到4095.0kg/hm2,新葵杂6号产量居第二位;除新葵杂4号和晋葵2号外,各品种具有较强的抗倒伏能力;而且从抗病性看,A15和新葵杂6号田间自然发病率低,抗性好.所以综合分析,A15和新葵杂6号两个油葵品种较适宜在当地盐碱地大力推广.     

Cytological Study of Callus and Regenerated Plants of Sunflower (Helianthus annuus L.)

In vitro culture of shoot apical meristems of Helianthus annuus was studied to determine the cytological condition of calli and regenerated shoots. A broad chromosome mosaicism (aneusomary) in bath callus and regenerated shoots was found, reflecting the cytological conditions of the explain. During in vitro plant development, a diplontic selection took place that was more rapid when very precocious in vitro flowering occurred.     

Plant Regeneration and Genetic Variability from Tissue Cultures of Sunflower (Helianthus annuus L.)

Adventitious buds were induced from in vitro culture of sunflower (Helianthus annuus L.) cotyledons. Four inbred lines (G1, G2, G3 and HA89), an open pollinated variety (‘Argentario’) and two hybrids with specific genetic markers were used. Cotyledons were cultured in vitro on MS medium (Murashlge and Skoog 1962) containing various concentrations of kinetin and indole 3-acetic acid (IAA). The quantitative interactions between auxin and cytokinin, the age of the cotyledons and the 2,3,5-triiodobenzoic acid (TIBA) treatments have been found to influence shoot regeneration. The plantlets, after rooting, were successfully established on soil. Qualitative variation was noted in self-pollinated progeny of plants regenerated from culture of two inbreds. Chimerism in regenerated plants was indicated by sectoring of the genetic markers. Some culture-induced variant phenotypes were similar to known spontaneous mutation in sunflower but others have been not yet described. Preliminary data indicate that most of them may have single-gene recessive inheritance.     

Genetic control of in vitro-organogenesis in recombinant inbred lines of sunflower (Helianthus annuus L.)

Ninety-three recombinant inbred lines (F₈) of sunflower developed by the single-seed descent method from the cross ‘PAC-2 × RHA-266’ were used to screen their regenerability by organogenesis. The experiment was designed in randomized complete blocks with three replications. Each replication consisted of 10 Petri dishes with four explants. Cotyledons were excised from 2-day-old seedlings. Each cotyledon was divided into two pieces (four explants), which were incubated in solid regeneration medium consisting of full-strength Murashige and Skoog medium modified by adding hormones. A high genotypic variability for organogenesis parameters between genotypes was observed in this study. The difference between all recombinant F₈ lines and their parents was not significant, showing that the 93 inbred lines used in this experiment are representative of the total possible recombinant lines from the cross ‘PAC-2 × RHA-266′. These F₈ lines were not consciously selected for any trait; therefore, they represent a random set of lines segregating for the organogenesis parameters, as well as for other traits which could be important for breeding.     

Origin and Fate of Aneusomaty in Sunflower (Helianthus annuus L.)

Chromosomal variability in Helianthus annuus was studied during the ontogenetic cycle of the plant. Aneusomaty originates during embryo development, progressively decreases in the course of plant growth and disappears at pre-meiosis. A number of aberrant mitoses with lagging chromosomes were found in all embryonic stages examined; by contrast, mitoses never showed aberrations in the shoot apices of plants.     

不同发育时期菊芋块茎DNA提取效果比较研究

为筛选菊芋块茎DNA提取的适宜生育时期,以“青芋1号”菊芋品种为试材,用改良CTAB法对不同发育时期菊芋块茎DNA进行提取,经琼脂糖凝胶电泳及全波长分光光度计检测总DNA的纯度、浓度及质量,选用4条ISSR引物进行PCR验证.结果表明:不同发育时期提取菊芋块茎DNA效果较好,DNA浓度呈现单峰曲线变化,峰值出现在第9周,与琼脂糖凝胶电泳检测呈现的亮度结果一致,PCR验证结果显示,提取的DNA能够满足后续相关分子生物学的要求.     

向日葵PMADS2 LIKE基因的克隆及表达分析

MADS-box家族基因是调控花发育进程的关键转录因子。在向日葵中大量的花发育相关基因有待于发掘和鉴定。本研究以向日葵(Helianthus annuus)为材料克隆到了一个MADS-box新成员PMADS2 LIKE。序列分析结果显示该基因具有MADS-box家族典型的MIKC保守结构域;系统发育树分析发现该基因与拟南芥PI基因亲缘关系最近;q RT-PCR组织表达模式分析该基因仅在花和果实中表达,且在6个花器官中仅在雄蕊和花瓣表达;q RT-PCR定量分析表示该基因从开花前25 d一直表达到开花期,并且在开花前5 d表达量最高。研究结果表明,PMADS2 LIKE基因在花发育和果实发育中具有一定的生物学功能。本研究的结果和推论为进一步探究向日葵PMADS2 LIKE基因在花发育和果实发育中的生物学功能提供前期数据基础,为理清MADS-box基因在向日葵花发育中的调控网络提供线索。     

菊芋营养器官解剖结构及其与抗旱性的关系

为了探讨菊芋适应干旱环境的解剖特征,分析菊芋植株营养器官解剖结构与抗旱性的关系,本研究采用石蜡切片和光学显微技术,对2个菊芋品种的根、地上茎、匍匐茎、叶的解剖结构进行了观察比较与参数测定。结果表明:‘青芋1号’与‘青芋2号’的根、地上茎、匍匐茎及叶的解剖结构均存在显著差异。与‘青芋2号’相比,‘青芋1号’根的表皮、皮层厚度分别显著减小35.21%、49.06%,内皮层、韧皮层厚度、韧皮部和木质部所占的比例分别是‘青芋2号’的1.23倍、1.33倍、1.34倍、1.57倍;‘青芋1号’地上茎和匍匐茎表皮的厚度分别是‘青芋2号’的1.18倍和1.38倍;在叶片的解剖结构上,‘青芋1号’的叶片厚度、上表皮、下表皮、栅栏组织厚度分别显著高于‘青芋2号’71.63μm、1.57μm、2.48μm、19.25μm。综合分析说明,‘青芋1号’通过根部表皮、皮层厚度的减小,内皮层、韧皮部的增厚及韧皮部和木质部所占比例的增加来响应干旱胁迫;通过地上茎和匍匐茎表皮厚度的增加,抵御水分流失;通过叶片、上表皮、下表皮的增厚及其内部栅栏组织的增多,利于水分的储存,从而适应干旱环境。本研究结果为菊芋抗旱性品种的鉴定提供解剖学依据。     

油葵HaDGAT2基因的功能及表达特性分析

二酰甘油酰基转移酶(DGAT)在植物油脂合成中起关键作用,其活性高低与植物含油量显著相关。为探明油葵二酰甘油酰基转移酶2基因(HaDGAT2)的功能,本研究以‘新葵杂5号’为试验材料,从油葵中扩增HaDGAT2基因序列并构建酵母穿梭表达载体pYES2-Ha DGAT2,将重组载体转入酿酒酵母INVSc1后提取酵母总脂肪酸,甲酯化后GC-MS分析。结果显示,在酿酒酵母中可成功诱导HaDGAT2基因,且转HaDGAT2基因酵母中棕榈油酸(16:1)和油酸(18:1)含量与对照相比得到提高。结果表明,HaDGAT2基因在调控油脂合成的过程中具有重要作用。实时荧光定量PCR分析结果表明HaDGAT2基因在油葵根、茎、叶、花、子叶和不同发育时期的种子中都有表达,且在开花后31天的种子中表达量高,说明该基因表达无组织特异性,在油葵种子油脂积累后期起关键调控作用。本研究为深入了解油葵油脂和调控机制提供了基础,为今后利用分子育种手段提高并改良葵花籽油的品质提供了理论基础。