水稻细胞质雄性不育育性回复株的基因型鉴定

通过对在套袋繁殖产生的滇Ⅰ型不育系合系42A群体中发现的可育株、及其与不育系杂交产生的F1和可育株自交S1代的育性和核恢复基因位点分析,发现不育系合系42A中出现的大约0.11%可育株包括两种类型。一类细胞质正常可育,核无恢复基因,基因型为N（rf/rf）,类似保持系,认为是保持系混杂所致。另一类可育株的细胞质雄性不育,核有恢复基因,其恢复基因位于水稻Rf-1基因区域,基因型为S（Rf/rf）。这类可育株不可能来自异品种或保持系串粉,可能是细胞核携带的育性相关基因发生育性自然回复突变导致。本研究将这类可育株简称为“育性回复株”。     

大白菜细胞核雄性不育两用系的利用途径探讨

不同来源的大白菜核不育两用系存在不同的遗传机制,其不育株与可育株都各略占50％并存。在利用上,50％不育株可作母本用于配制F_1杂种和核基因互作雄性不育系;50％可育株可用作F_1杂种父本、质核复合雄性不育保持系和核基因互作雄性不育临保系,同时自交还可快速选育自交系。在优势育种上,大白菜核不育两用系具有一定开发潜力和应用前景。     

核不育复等位基因向娃娃菜中转育的研究

根据"复等位基因遗传假说",以甲型两用系AB12为不育源(直筒生态型),采用杂交、自交、兄妹交的方法,将核不育基因向娃娃菜可育品系06006中转育,获得了不育株率100%的新核基因雄性不育系,扩大了原有不育系的遗传基础,拓宽了该优良雄性不育基因的应用范围.     

核不育油用亚麻研究初报

1975年,于本院亚麻试验田里发现一株不育油用亚麻,经鉴定是一株无花粉型的、细胞质正常的、受一个显性雄性不育单基因(Ms)控制的天然突变体,在我国尚系首次发现.它的花为淡紫色、种皮近似白色.不育株与可育株杂交后,F_1代分离出来的不育株与可育株的比例为1:1;其中可育株自交后代育性不分离;不育株的育性分离没有中间类型,不是全育,就是全不育.这种核不育亚麻是进行亚麻育种的良好杂交材料.     

谷子(Setaria italica)显性雄性不育基因的发现

1978年我们在“澳大利亚谷×吐鲁番谷”的杂交后代中,得到1份雄性不育材料,经7个遗传世代的研究发现,其不育株测交和回交,后代育性始终按1∶1分离,自交后代育性分离为3(不育)∶1(可育);其可育株育性不分离。它的不育性是受核内的显性雄性不育基因控制的。这是在谷子中的首次发现。它在理论研究、杂种优势利用和谷子育种中都有很高的价值。     

谷子不育性的化学保持研究

以谷子高度雄性不育系927A和其近等基因可育系927C为材料,研究了不育株与可育株内源激素含量水平的差异,结果表明927A的败育与低水平的内源细胞生长素(IAA)有关.对927A施用外源细胞生长素(IAA)可部分恢复其雄配子的育性,显著提高其当代的自交结实率,证明IAA对927A的不育性具有化学保持作用.     

黏类小麦细胞质雄性不育相关基因cMDH的克隆与表达分析

为深入研究黏类小麦雄性不育的分子遗传机制,利用抑制差减杂交技术构建了黏类小麦育性相关基因的二核期SSH文库.经文库筛选后,得到一个在可育文库中表达的与胞质苹果酸脱氢酶基因同源的EST序列.以该EST序列为信息探针,经电子克隆、RT-PCR、PCR克隆与序列分析,获得了小麦胞质苹果酸脱氢酶(cytosolic malate dehydrogenases,cMDH)基因的cDNA与DNA序列,利用荧光定量PCR技术对该基因在不育株和可育株花药中的表达进行了分析,并比较了MDH在小麦不育株和可育株中的活性变化.结果表明,该基因的cDNA序列长1213 bp,编码333个氨基酸;DNA序列长2 908 bp,含有7个外显子和6个内含子;该基因在不育株和可育株花药发育3个时期(单核、二核和三核)的表达均表现为先升后降的模式,而且该基因在不育株花药发育的二核期和三核期的表达相对于可育株被明显抑制;MDH在小麦不育株和可育株中的活性变化趋势与定量结果一致.推测该基因在花粉发育早期表达,它的下调表达可能影响了小麦雄蕊发育过程中的能量供应而导致雄性不育.     

棉花核不育系豫98-8A育性遗传分析

为了阐明1999年从转基因后代遗传群体中发现的1株雄性不育植株不育基因的遗传规律及其与现有不育基因的等位性,采用表型观察测量,以及经典的自交和测交手段,研究了该不育材料败育性状的遗传规律。花器官形态特征调查表明：不育株花柱长和花柱外露长度均明显高于同质系的正常可育株,而每朵花的子房直径及花药数量没有明显差异。遗传分析表明：杂合体可育株自交,后代不育株与可育株呈3:1分离,不育株与杂合姊妹可育株测交,不育株与可育株呈1:1分离,表明该核不育材料受隐性单位点控制;与阆A(msc1)、洞A(msc3)等育性位点杂合可育株分别杂交,其F1代单株育性均得到恢复。由其F1代产生的F1:2家系中均出现不育株与可育株呈1:3和7:9两个育性分离群体,表明该材料败育基因为不同于阆A、洞A的不育基因位点。     

甘蓝型油菜轮回选择研究

以同时携带显性核不育基因Ms和显性不育抑制基因Rf的甘蓝型油菜为父本,含有相应隐性等位基因的同类型油菜为母本,经一代杂交成功地把显性核不育基因导入具不同细胞质来源的基因型中.90%以上的杂种一代\回交和复交组合均表现为1可育:1不育的分离.在第一批转育所用的15个亲本中,有3个对由Ms基因控制的雄性不育性有抑制作用,它们     

食荚豌豆雄性不育突变体的遗传研究

对国内首例豌豆雄性不育突变体的不育度、遗传特点及稳定性进行研究。观察发现：在生育前期。不育株外部形态特征与正常株没有明显差异;现蕾后,剥开花蕾可看到不育株的花药呈淡黄色半透明状。而可育株的花药呈橙黄色。用I2-KI染色法镜检花粉的可染性,发现不育株的花药内没有花粉粒,败育彻底,为典型的“无花粉型”雄性不育。用不育株作母本,与同品系的正常可育株进行姊妹交,F1全部可育,F2可育株与不育株的分离比例为3：1。用不育株作母本,其他品系作父本进行测交,同时用其他品系作母本,姊妹交F1作父本进行反交,正反交后代的育性表现一致。F1全部可育。F2可育株与不育株呈3：1分离。结果表明：该雄性不育突变体的不育性是可遗传的,属单隐性基因控制的核不育类型,与细胞质遗传物质无关。在不同年份、不同季节下,不育性状表现稳定。     

Pollen-mediated gene flow in an autogamous crop: Foxtail millet (Setavia italica)

Gene How is a key concern associated with the eonlamination of seed multiplication iields and the use of transgenic crops. The release of herbicide-resistant germplasnis and the use of male-sterile varieties make foxtail millet (Sviaria ilalicii) an appropriate material to inves-tigate this concern. Pollen dispersal from pollen donor sources and gene tlow in fertile and male-slerile varieties of fortail millet were investigated in experiments in China and France. Although foxtail millel is mainly autogamous (less than 2% oiitcrossing), pollen from a 5m diameter plot could fertilize a tertile variety up to 24 ni away, and a male-sterile variety up to 40m. Negative exponential and WeibuU fnnctions were used to tit the relationships between percentage and number i.A iubrids per cm of ear with distance from the pollen source. Wind velocity and direction resulted in variations of gene How intensity, but did nol change liie slope ofthe regression equations. Pollen competition for available target ovules was observed up to 2 m away from a 1 m diameter pollen source. The percentage of pollen grains that fertilized ovules outside the pollen donor source was 1.4% of the total pollen grain produetion. For every iflO pollen grains released under lieid conditions 1.5 seeds were produced. The probability funetion of pollen dispersal for one donor plant was calculated. It allowed predictions of gene How accord-ing to lield shapes, estimations of isolation distances to preserve /ariety purity against pollen contamination, and the fornuiiation of efficient male/female ratios Ibr hybrid seed production.     

Investigation of pollen competition between wild and cultivated sorghums (<Emphasis Type="Italic">Sorghum bicolor</Emphasis> (L.) Moench) using simple sequence repeats markers

In self-compatible plant species stigmata receive a mixture of self and outcrossed pollen and competition between them is expected to play a major role in determining the pollen-mediated gene flow. The use of male sterile bait plants in field trials to demonstrate the rate of gene flow is questionable due to lack of pollination competition. However, little direct evidence has been published. A field experiment of male sterile and male fertile sorghum pollen recipient bait plants was conducted to evaluate pollen competition between wild and cultivated sorghums and the effects of pollen competition on gene flow assessment. Pollen competition between wild and cultivated sorghums was estimated from two-component pollen mixtures of wild and cultivated sorghum (1:1 ratio) applied to wild, cultivated and male-sterile maternal bait plants. Paternity was determined in the progeny using two diagnostic Simple Sequence Repeat markers. The study found that self pollen has higher seed-siring success. Maternal genotype influences the siring ability of the pollen donor components which significantly deviated from the 1:1 pollen loads. The study showed that published estimates of gene flow derived from studies using male-sterile bait plants seriously overestimate gene flow and that pollen competition may be a significant factor influencing outcrossing rates. The results suggest that the predominant direction of gene flow is from cultivated to wild sorghum, potentially leading to introgression of crop genes into wild sorghum. Pollen competition should be taken into account in gene flow estimation, since presence of self-pollen can account for over half of seed produced irrespective of maternal genotype.     

Induction of an alloplasmic male-sterile Brassica oleracea by substituting cytoplasm from ‘Early Scarlet Globe’ radish (Raphanus sativus)

Summary Alloplasmic male-sterile Brassica oleracea L. was synthesized in a backcrossing program through amphidiploid Raphanobrassica by using Early Scarlet Globe radish (Raphanus sativus L.) as the donor of cytoplasm and B. oleracea broccoli and cabbage as recurrent pollen parents. Persistence of radish chromosomes and high female sterility were encountered in the first four backcrosses. Following use of colchiploid 4x broccoli as pollen parent, a BC₅ plant was obtained that had 2n=3x+1=28 chromosomes, improved seed set, and no radish traits. The BC₆ with recurrent 2x broccoli contained male-sterile plants with 2n=18 or 19 chromosomes, increased seed set, and broccoli morphology. Subsequent generations segregated for male-sterile and restored male-fertile plants, some with variable development of stamens and pollen. Leaf color of the alloplasmic plants, especially seedlings, was lighter green than normal.     

Restrictions of the efficiency of haploid plant production in apple cultivar Idared,through parthenogenesisin situ

Summary For haploid plant production in Idared apple, parthenogenesisin situ was induced by irradiated pollen. The most important restrictions were seed set, embryo germination and green plant recovery from germinated embryos. The efficiency of green plant production was strongly influenced by irradiation dose, picking time and quality of the irradiated pollen. 250 Gy pollen was more efficient for green plant production than 500 Gy pollen. Using 250 Gy pollen with reduced germination capacity resulted in a five-fold lower green plant yield. Delaying picking of the fruits (140 instead of 70 days after pollination) had a significant positive effect. Up to 20 plants per 1000 pollinated flowers, showing no expression of a marker gene present in the pollen parent, were obtained in some treatments. Induction efficiency was high when seed weight was high. This offers the possibility of selecting seeds with the best chances for haploid plant production. Heavy seeds generally contained visual normal developed embryos, which germinated better than those from lighter seeds and gave normal plant development. Delayed picking increased the proportion of large seeds, suggesting that pollination with irradiated pollen retards embryo development. From treatments with irradiated pollen of poor quality more small seeds were harvested.     

‘砀山酥梨’花柱可授性及蜜蜂授粉特性的研究

采用半离体培养法和联苯胺-过氧化氢法研究了‘砀山酥梨’花粉萌发特性和柱头可授性。通过对‘酥梨’的最适授粉时间以及7种授粉处理坐果率的研究,探讨其授粉特性。结果表明：开花后每天酥梨柱头活性最佳的时间是11时至15时,柱头最佳授粉时间为开花后第4~5天;‘酥梨’花粉最适萌发温度为25℃,花粉管生长最有利的温度为22~25℃,在相同温度下,花粉萌发率和花粉管长度随时间增加而增加。7种授粉处理‘酥梨’的花朵坐果率中,没有昆虫授粉的‘酥梨’坐果率为0.48%和0.44%,自然授粉和人工授粉坐果率分别为2.83%和13.05%,‘酥梨’ב雪花梨’组蜜蜂授粉的坐果率为32.95%,新鲜花粉和商品花粉坐果率分别为70.2%和13.05%。由此可见,‘酥梨’是异花虫媒果树,蜜蜂可以在‘酥梨’柱头活性最高时进行授粉,从而提高‘酥梨’的坐果率,同时通过对‘酥梨’花器官生理活性和坐果习性的研究,为‘酥梨’的授粉和生产提供基础资料。     

Study of morpho-physiological, phenological and reproductive behaviour in protogynous lines of Indian mustard [Brassica juncea (L.)]

Understanding the relationship between floral and reproductive traits are critical to understand the evolution of plant species in brassica and need for selecting lines as parental lines in hybrid seed prodcution. The protogyny nature of Brassica species are characterized based on the variations in floral morphology, protogyny interval, stigma receptivity, pollen biology, compatibility and seeds set. The variation in stigma morphology features (stigma exsertion prior to anthesis) and protogyny interval (ranged from 8 to 10 days) formed phenotypically distinct. Studies on stigma receptivity and pollen viability revealed significant variability and found to be at its peak up to 3 days after anthesis and reduced drastically thereafter. Compatibility crosses and post pollination events such as pollen deposition on the surface of the stigma and culminating with the entry of the pollen tube into the embryo sac, was rapid and completed within 2 days after pollination, whereas in in-compatible cross not even single pollen was germinated on stigmatic surface and consequently no pollen tube growth even after sixth day of pollination. In compatible crosses average seed set ranged from 3 to 8 seeds per pod from cross pollination and no seed set in self. The investigation revealed that variation exists for protogyny interval, stigma receptivity and compatibility in the protogynous lines of Indian mustard and this can be exploited for the production of hybrids without emasculation.     

Reproductive barriers in the intergeneric hybridization between <Emphasis Type="Italic">Chrysanthemum grandiflorum</Emphasis> (Ramat.) Kitam. and <Emphasis Type="Italic">Ajania przewalskii</Emphasis> Poljak. (Asteraceae)

In order to reveal the causes of low seed set in the intergeneric cross between Chrysanthemum grandiflorum (Ramat.) Kitam. ‘Zhongshanjingui’ (female parent) and Ajania przewalskii Poljak. (male parent), we systematically investigated megasporogenesis and megagametogenesis of female parents, pollen germinability of male parents, pollen–pistil interaction and hybrid embryo development using paraffin section and light, fluorescence and scanning electron microscopy. The results indicate that development of megagametophyte of female parent belongs to Polygonum type and 84.6% ovules developed to normal 8-nucleate embryo sac at the stage of pollination; the pollen germinability in vitro of male parent was 39.3%; a large number of pollen grains germinated normally and no callose deposited on the female stigmas within 24 h after pollination. Furthermore, about 75% ovules were fertilized, whereas only 17.2% ovules developed to normal maturing embryos at 15 days after pollination due to abortion at various developmental stages. Taken together, these results suggest that sterile ovules of female parent, pollen viability and pollen–pistil interaction have no significant influence on seed set in the cross. A high percentage of embryo abortion or degeneration, i.e. post-fertilization barrier, may be the main factor for low seed yield.     

华北驼绒藜花粉萌发特性及不同授粉方式对其结实率的影响

为寻找华北驼绒藜结实率低、群落退化的原因,在栽培条件下对其花粉生活力、寿命、柱头萌发率与结实率的关系进行了相关性研究。采用同株、异株、自然授粉处理华北驼绒藜,同株授粉结实率(81.53%)显著低于异株授粉(88.6%)和自然授粉(92.6%)的结实率;采用TTC法对华北驼绒藜花粉寿命和生活力的研究结果表明,其平均寿命为6d,最佳授粉时间为1~3d,生活力达95%左右,不存在花粉败育和短命的问题,即不存在小孢子发育障碍。3种授粉方式处理后,采用荧光显微镜观察华北驼绒藜花粉在柱头上的萌发率,萌发高峰集中在授粉后3~4h,萌发率达90%以上,三者不存在显著差异,即华北驼绒藜不存在花粉与柱头的识别障碍。结实率与花粉在柱头萌发率的相关分析结果表明,二者不存在相关性。研究初步认为,同株授粉产生结实率低的原因可能发生在授精或胚胎发育过程,有待进一步研究。目前,华北驼绒藜野生群落严重退化,植被密度减少,可能存在大量自株授粉是造成群落退化的原因之一。     

Floral morphology affects seed productivity through pollination efficiency in radish (<Emphasis Type="Italic">Raphanus sativus</Emphasis> L.)

To examine the effect of stigma position and size on seed productivity through pollination efficiency in radish, the numbers of self and cross pollen grains on the stigmas and the seed productivity under insect-pollination were compared among four bred lines. Lines with a small stigma or a high stigma relative to the anthers on long stamens tended to receive fewer self and total (self + cross) pollen grains but showed a higher cross-pollination percentage (ratio of cross pollen grains to total pollen grains on stigmas) than those with a large stigma or a lower stigma. Additionally, a higher cross-pollination percentage was associated with a higher outcrossing percentage. This result suggests that variations in the two stigma traits affect the outcrossing percentage via the cross-pollination percentage. Therefore, it should be possible to prevent loss of F₁ purity by selecting new parental lines with floral morphology that favors cross-pollination percentage.     

Rapid method of determining pollen incompatibility in Chrysanthemum morifolium Ramat

Summary A classification system for predicting high, medium and low seed set from Chrysanthemum morifolium crosses was developed. Stigmas collected and preserved 8 days after pollination were examined with a binocular microscope for adhering pollen grains. Florets from compatible crosses had brown stigmas with pollen grains attached and embryo development while florets from incompatible crosses had white stigmas with no or rarely 1 or 2 pollen grains attached and no embryo development. Using this method the occurrence of seed set was predicted with 78.5% accuracy in one year and 89% in the following. This provides a method of determining the compatibility of a chrysanthemum cross without waiting the 1 to 2 months required for seed ripening.Scientific Journal Series Paper Number 7570 of the Minnesota Agricultural Experiment Station.