不同倍性栽培稻与药用野生稻种间杂交不结实性的比较胚胎学观察

以栽培稻(Oryza sativa)品种L202二倍体（L202 2x）及其同源四倍体（L202 4x）作母本分别与药用野生稻（Oryza officinalis）杂交,比较了两个杂交组合的杂交结实性及影响结实性的杂种胚胎发育细胞学。两个杂交组合所取得的一致结果是：杂交授粉后30 d所有杂交小穗均干枯死亡,不能结实;种间杂交不结实的原因在于受精和杂种胚胎胚乳发育异常。两个杂交组合的受精和杂种胚胎胚乳发育虽然均存在双受精、单受精和未受精类型,但就受精率而言,L202 2x作母本的平均总受精率为59.45％,略高于L202 4x（54.87％）; L202 4x的单受精率（24.60％）则高于L202 2x的单受精率（15.30％）。两个杂交组合的杂种胚胎胚乳发育均严重异常,虽然在授粉后1 d形成小的球形胚;到授粉后3 d,胚胎发育停滞并开始解体;至授粉后5 d,只有二倍体杂交组合的极少数胚胎可发育到梨形胚时期,四倍体组合的胚胎则基本全部解体。双受精胚囊内胚乳发育更为异常,在授粉后1 d,游离胚乳核就开始解体退化,到授粉后3 d不能形成细胞化胚乳或细胞化过程异常。与L202 2x作母本的杂交相比,以L202 4x作母本所得杂种胚胎发育更为滞后,且退化严重,形成的游离胚乳核少,解体早,不能进行细胞化。因此,利用同源四倍体栽培稻与药用野生稻杂交获得种间杂种的难度更大。     

栽培稻与药用野生稻杂种后代的形态学和细胞遗传学研究

 选用两个抗褐飞虱的药用野生稻编号材料与两个感虫栽培品种杂交，通过胚培养获得了F₁、BC₁F₁和F₂植株。F₁杂种表现出野生亲本紫色柱头、长芒及抗褐飞虱等特性，其花粉母细胞中期I染色体多为单价体(仅0.77个二价体)，甚至不能形成花药，因而表现雄性不育，仅获得一株F₂，染色体数目为2n=46。用栽培稻回交获得BC₁F₁植株为异源三倍体，中期I染色体构型为12.25Ⅰ+11.85Ⅱ+0.01Ⅲ。     

稻属种间天然异交种的分子细胞学鉴定

利用基因组荧光原位杂交（GISH）技术快速鉴定了栽培稻与野生稻的天然异交种的基因组组成,分析了该杂种在减数分裂中期Ⅰ的染色体配对情况。根据根尖细胞的染色体数目,发现该杂种是具有36条染色体的三倍体;通过减数分裂中期Ⅰ染色体的配对研究,发现该杂种染色体很少发生配对,绝大部分染色体以单价体形式存在;结合GISH技术的分析,证实该杂种是由A、B和C 3个染色体组组成。因此该杂种是栽培稻和小粒野生稻的天然杂交种。     

Progress on Transferring Elite Genes from Non-AA Genome Wild Rice into Oryza sativa through Interspecific Hybridization

The progress of research on transferring elite genes from non-AA genome wild rice into Oryza sativa through interspecific hybridization are in three respects, that is, breeding monosomic alien addition lines (MAALs), constructing introgression lines (ILs) and analyzing the heredity of the characters and mapping the related genes. There are serious reproductive barriers, mainly incrossability and hybrid sterility, in the interspecific hybridization of O. sativa with non-AA genome wild rice. These are the ‘bo...     

利用种间杂交途径向栽培稻转移非AA组野生稻有利基因的研究进展

利用种间有性杂交向栽培稻转移非AA组野生稻有利基因的研究在三个方面取得进展,即单体异附加系的培育、渗入系的建立以及有利性状的遗传及基因定位。栽培稻与非AA组野生稻种间杂交存在严重的生殖障碍,表现在不可交配性和杂种不育性两个方面,这是限制非AA组野生稻有利基因转移的“瓶颈”。传统的有性杂交手段和生物技术相结合是克服种间生殖障碍,提高非AA组野生稻有利基因育种利用效率的可行途径。     

同源四倍体水稻与非洲栽培稻杂交的胚胎学观察

以同源四倍体水稻(Oryza sativa,2n=4x=48)和二倍体水稻(Oryza Sativa,2n=2x=24)为母本,以非洲栽培稻(Dryza glarri,2n=2x=24)为父本进行远缘杂交配组,对其杂交结实的胚胎学机理进行了研究。结果表明,非洲栽培稻的花粉粒能在二倍体水稻的柱头上萌发,但萌发频率仅为0．18％～0．56％,花粉管在花柱内的伸长速度比较慢,发生受精的频率比较低,因而二倍体水稻与非洲栽培稻杂交的结实率比较低。然而,非洲栽培稻的花粉粒在同源四倍体水稻的柱头上萌发频率高达12．81％～14．65％,花粉管在花柱内的伸长速度比较快,发生受精的频率比较高,因而同源四倍体水稻与非洲栽培稻杂交的结实率比较高。在同源四倍体水稻与非洲栽培稻的杂交中发现,其受精作用比较特殊,其中包括精细胞与卵细胞的单受精作用,精细胞与次生极核的单受精作用和正常的双受精作用,总受精率和总成胚率分别为7．49％～9．28％和6．61％～8．64％。     

栽培稻种间近等基因系杂种育性研究

 为了研究亚洲栽培稻与非洲栽培稻种间杂种不育的遗传基础，以亚洲栽培稻品种WAB56-104为测验种与来自种间回交组合WAB56-104/CG14//WAB56-104///WAB56-104的14个种间近等基因系测交，对亲本及杂种的花粉及小穗育性研究表明，这14个近等基因系带有来自非洲栽培稻品种CG14的3个互不等位的配子消除位点和2个互不等位的花粉不育基因位点, 其中至少有2个配子消除位点未曾报道过。     

栽培稻种间近等基因系杂种育性遗传及其基因定位

 用粳型亚洲栽培稻品种WAB56-104与来自种间回交组合WAB56-/104/CG14∥WAB56-104///WAB56-104的3个栽培稻种间近等基因系杂交、回交，对3个BC1F1群体的花粉及小穗育性遗传研究表明，育性遗传符合单位点孢子体-配子体互作模型，来自非洲栽培稻的不育基因在与相同位点的亚洲栽培稻等位基因互作时，导致携带亚洲栽培稻等位基因的雌雄配子败育，而形成花粉及小穗的半不育，但本研究的3个组合中，并未导致所有携带亚洲栽培稻等位基因的雌配子完全败育，其作用介于配子消除与花粉灭杀之间。用微卫星标记对这3个群体的育性基因定位表明，它们都位于第6染色体短臂末端，但WAB450-2、WAB450-8携带的不育位点与紧密连锁的微卫星标记RM190、RM133共分离，且很可能与S1等位，而WAB450-7的不育基因则与位于相邻区间的RM253呈松散连锁，可能与WAB450-2、WAB450-8的育性位点不同。     

Molecular Cytogenetic Analysis of Spontaneous Interspecific Hybrid Between Oryza sativa and Oryza minuta

Genomic in situ hybridization (GISH) is a powerful tool to characterize parental chromosomes in interspecific hybrids, including the behaviour of autosynapsis and chromosome paidng. It was used to distinguish the chromosomes of Oryza sativa from wild species in a spontaneous interspecific hybdd and to investigate the chromosome pairing at metaphase I in meiosis of the hybdd in this study. The hybrid was a triploid with 36 chromosomes according to the chromosome nurnber investigated in mitosis of root tips. During metaphase I of meiosis in the hybrid, less chromosome pairing was observed and most of the chromosomes existed as univalent. Based on GISH and FISH (Fluorescent in situ hybridization) analyses, the chromosomes of the hybrid were composed of genomes A, B and C. Thus, it was believed that the hybrid was the result of natural hybridization between cultivated rice and wild species O. minuta which was planted in experimental fields.     

广东高州普通野生稻与粳稻杂交F1胚囊育性及发育特点

利用整体染色激光扫描共聚焦显微术(WE CLSM), 对广东高州普通野生稻与粳稻台中65杂交F1的成熟胚囊育性、胚囊形成发育及受精特点进行研究。杂种F1成熟胚囊平均育性为64.61%。不同杂种的成熟胚囊均存在不同程度的异常现象, 异常的主要类型有：胚囊退化、小胚囊、雌性生殖单位退化、卵器退化、极核数目或位置异常及其他异常等类型。其中频率最高的异常类型为雌性生殖单位退化（10.49%）,其次是胚囊退化（9.80%）。杂种F1的胚囊发育过程与高州野生稻原种基本一致,育性低的杂种F1胚囊发育过程出现多种异常现象。对3个杂种F1受粉后1 d的胚囊进行观察发现,除正常受精膨大的胚囊外,还存在结构正常未受精和结构异常未受精等异常现象。受粉后7 d的胚囊基本上与受粉后1 d的相似。研究结果表明,广东高州普通野生稻可能具有决定杂种F1胚囊育性的不同基因,包括中性基因等。     

我国三种野生稻及其与栽培稻种间杂种外稃表面显微特征的比较观察

 用扫描电镜对栽培稻的两个籼型品种（Oryza sativa subsp. indica）、普通野生稻（O．rufipogon）、药用野生稻（O. officinalis）、疣粒野生稻（O. meyeriana）及野生稻与栽培稻的种间杂种F₁的外稃表面显微特征进行了比较研究,观察到外稃表面的显微特征有明显的种间差异,并初步概述了三种野生稻的乳突特征。栽培稻与野生稻的种间杂种外稃显微特征受亲本影响,其中乳突座间距、列宽主要受野生稻亲本影响;底座以上的峰突特征如峰距、峰数则表现为栽培稻亲本的特征;乳突分布密度表现为超亲现象。     

Grain Protein Content of Interspecific Progenies Derivedfrom the Cross of African Rice (Oryza glaberrima Steud.) and Asian Rice (Oryza sativa L.)

Abstract

The protein contents of the grain of 50 interspecific progenies developed from the cross between WAB56-104, an Oryza sativa variety, and CG 14, an Oryza glaberrima line, were investigated. In contrast to the higher protein content of O. glaberrima than O. sativa on the average, the protein content of CG 14 was always lower than that of WAB56-104. However, judging from the average of three seasons, 72% of the interspecific progenies had a higher protein content than the mid-parent and 50% of them had a higher protein content than WAB56-104. Although the actual values of protein content of the interspecific progenies were significantly different among the seasons, a highly significant correlation was always observed in protein content between any two of the three seasons. Protein content therefore was considered character of each interspecific progeny though it was also affected by environment. A significant correlation was not observed between paddy yield and protein content in any season; several interspecific progenies showed higher protein content and paddy yield than the mid-parents. A low paddy yield is likely to be associated with high protein content through physiological regulation without a genetic linkage between the two traits. However, the results suggest that the transgressive segregation of protein content observed in the interspecific progenies is attributed not to this physiological regulation but to a certain mechanism to concentrate protein in grains with a genetic background.     

同源四倍体水稻与非洲栽培稻杂交的效果

以4份同源四倍体水稻(Oryza sativa,2n=4x=48)和4份二倍体水稻(Oryza sativa,2n=2x=24)为母本,以4份非洲栽培稻(Oryza glaberrima,2n=2x=24)为父本进行远缘杂交配组,共配制32个杂交组合。研究结果表明,非洲栽培稻分别与亚洲栽培稻中的两种倍性水平的水稻品系进行杂交均表现出一定的可交配性,生殖隔离并不很严格。然而,不同倍性的亚洲栽培稻与非洲栽培稻杂交,其结实率表现出明显的差异。二倍体水稻与非洲栽培稻不容易杂交,其结实率比较低(0．32％—1．93％),而同源四倍体水稻与非洲栽培稻较容易杂交,其结实率比较高(1．22％—8．86％,)。由此认为,利用同源四倍体水稻为母本与非洲栽培稻杂交更容易将后者的遗传物质引入亚洲栽培稻。     

水稻 (Oryza sativa L.)花粉及花药壁发育的超微结构研究

 运用透射电子显微镜技术,系统观察了水稻花粉及其花药壁层的发育过程,发现了一些新的现象：(1)小孢子母细胞减数分裂期间,伴随核内染色体变化的同时细胞质发生了“改组”现象,主要表现为核糖体分布密度的规律性变化,这标志着孢子体向配子体的转变。(2) 小孢子外壁的发育始于四分体晚期,最早表现在四分孢子质膜上沉积了少量的壁物质。随后沉积增多,至小孢子早期即形成初生外壁。此后外壁发育迅速,到小孢子晚期外壁已基本发育完成。(3)小孢子中期,小孢子细胞核的双层核膜局部分开,并逐渐扩张成一个“大泡”。核膜扩张在这一时期是一种普遍现象。(4)在花粉发育过程中,绒毡层细胞结构发生明显变化：小孢子母细胞形成之初,绒毡层细胞结构完整,内质网极少;随着减数分裂的进行,绒毡层胞质浓缩,细胞内出现“空腔”,内质网丰富;到了小孢子中期,仍有较多堆叠的内质网,此后逐渐消失。表明内质网在绒毡层的发育中起着重要的物质合成及运输作用。(5)花粉完全成熟时,花药中层细胞的壁以及绒毡层的外切向壁紧贴在一起,形成了一叠合的“壁”结构。     

Natural Homologous Triploidization and DNA Methylation in SARII-628, a Twin-seedling Line of Rice （Oryza sativa）

A total of five pairs of diploid-triploid twin-seedlings (a diploid seedling and a triploid seedling emerged from a grain) were selected out from 4500 pairs of seedlings from SARII-628, a twin-seedling rice line. SSR analysis indicated that no difference between the diploid seedling and corresponding triploid seedling in a twin-seedling was found at the 310 loci, indicating that there was no obvious change in DNA primary structure. A modified AFLP technique ‘MSAP (methylation-sensitive AFLP)’ was used to analyze methylation mutation. Although no methylation mutation was noted among the five diploids, 29 methylation mutation loci were found from the corresponding triploids. This suggested that methylation mutation happened rapidly on M0 generation after natural homologous triploidization. The mutations were classified into 10 types, including 3 increased types, 3 decreased types and 4 undecided types of methylation-degrees. The bands of 22 loci were sequenced and then those sequences were searched through website. The result showed that the methylation mutation involved into the whole rice genome and the 12 pairs of chromosomes. The mutation trend was site-related and there were different mutation loci for different triploids, which foretold that SARII-628 would have different evolution fates after natural homologous triploidization.     

云南药用野生稻幼穗离体培养研究

药用野生稻由于结实率低、落粒性强,难以利用种子保存繁殖。为了进一步在育种中有效地利用药用野生稻抗虫、抗病、抗旱、分蘖力强等优良特性,本研究利用药用野生稻幼穗,通过离体培养方法获得植株再生苗。实验设计了10种不同生长调节剂浓度的愈伤诱导培养基,筛选出N6+2,4-D 2.0 mg/L+植物凝胶2.8 g/L+蔗糖30 g/L、pH值5.8是最适合愈伤诱导的培养基;设计了10种分化培养基,筛选出N6+6-BA 0.3 mg/L+NAA 0.3 mg/L+ZT 2.0mg/L+KT 3.0 mg/L+植物凝胶2.8 g/L+蔗糖30.0 g/L、pH值5.8最适合作分化的培养基。设计了10种生根培养基,1/2N6+NAA 0.2 mg/L+植物凝胶2.8 g/L+蔗糖15.0 g/L、pH值5.8最适合作生根培养基。研究发现用4℃低温对幼穗前处理,然后使用强弱光照交替进行预培养后再进行愈伤诱导,可获得较高的愈伤诱导率。     

同源四倍体水稻与非洲栽培稻杂交的后效性研究

利用亚洲栽培稻中的4份同源四倍体水稻(O.sativa,2n=4x=48)和相应的4份二倍体水稻(O.sativa,2n=2x=24)为母本,以4份非洲栽培稻(O.glaberrima,2n=2x=24)为花粉供体进行远缘杂交后,对其杂种后代的分离动态进行了研究.结果表明,不同倍性的普通栽培稻与非洲栽培稻之间杂交后代的结实率以二倍体普通栽培稻的较高.在配制的32个杂交组合中,其杂种第1代群体均表现出明显的营养生长优势.从群体的生长势来看,杂种第2代群体比杂种第1代群体要弱一些;在杂种第2代群体中,以同源四倍体水稻为母本的杂交组合的分离现象比以二倍体水稻为母本的杂交组合的分离现象更明显.在各杂交组合的第3代群体中,从植株的株叶形态和生育期来看,株系间的差异和株系内的变异依然很明显,变异频率更宽,变异种类更多.在普通栽培稻与非洲栽培稻的杂交组合中,育性变异、生长势变异、株叶形态变异、染色体变异和结实性变异等是较易发现的变异现象.     

根表铁胶膜对镉污染土壤水稻吸镉量的影响(英文)

通过采用镉污染的土壤—河砂—营养液联合培养的方法,研究了在水稻分蘖期施铁对水稻根表铁胶膜中铁及铁胶膜吸附镉的影响。设四个亚铁(Fe~(2+))处理水平分别为:0、30、50、70mg/L,研究施铁浓度与水稻根表铁氧化物胶膜的数量和吸附镉量之间的关系。结果表明,施铁处理的铁胶膜中铁的含量比不施铁(0处理)的高,但在其他三种处理 (30、 50、70mg/L)中,随施铁浓度的升高而降低。另外,施铁浓度与铁胶膜的数量呈正相关,所以,铁胶膜吸附镉与施铁浓度呈正相关。在水稻的另外三个时期:拔节、孕穗、灌浆期,对该问题将作进一步研究。     

Relationship between Shoot Elongation and Dry Matter Weight During Submergence in Oryza sativa L. and O. glaberrima Steud. Rice Cultivars

Abstract

Rice plants are damaged by flash floods with a rapid increase in water level caused by a heavy rain. However, rice plants cope with the flash floods either by an “escape strategy” involving rapid shoot elongation or by a “quiescence strategy” involving surviving under water with minimal activity. As we found in previous experiments, Saligbeli cultivar adapted well to flash floods through rapid shoot elongation. To understand the vigorous growth process during submergence, we studied the relationship between shoot elongation and changes in dry matter weight (DMW) during submergence. O. glaberrima Steud. cv. Saligbeli and O. sativa L. cv. Ballawé and IR 49830-7-1-2-2 were used. Saligbeli and Ballawé exhibit shoot elongation, and IR 49830-7-1-2-2 exhibits flash-flood tolerance due to the presence of the Sub-1 gene. Twelve-day-old seedlings were submerged for 7 days and the plant length and DMW were measured. The plant length ratio of submerged to control plants in Saligbeli was higher than that of other cultivars during 2-6 days of submergence but IR 49830-7-1-2-2 shoot elongation was inhibited by submergence. In all three cultivars, the elongation of the developing leaf sheath conferred shoot elongation during submergence. The plant length of all submerged plants showed a strong positive correlation with DMW of the leaves developed during submergence. Submerged Saligbeli and Ballawé showed strong negative correlations between DMW of the leaves developed before and during submergence (r = –0.786 for Saligbeli and –0.772 for Ballawé , P < 0.05), suggesting that the enhancement of shoot elongation during submergence is accomplished by using dry matter of the leaves developed before submergence. However, the correlation was not observed in the submerged IR 49830-7-1-2-2. Further details from studies using isotopes are also needed to understand the plant growth during submergence.     

栽培稻与普通野生稻BC2F2群体产量相关性状的QTL分析

以广陆矮4号(Oryza sativa ssp. indica)为母本及轮回亲本,普通野生稻（Oryza rufipogon）为父本,分单株连续回交2次,构建BC2F2群体。首先用241对具有双亲多态性的SSR标记对BC2F1单株进行代换片段分析,在此基础上,根据BC2F1的表型选产量较优的单株自交获得BC2F2群体,用代换片段上具有双亲杂合型基因型的24对SSR标记进行QTL定位。在所选的BC2F1单株上,共检测到分布于7条染色体上的20个野生稻的代换片段,平均每条染色体上有2.86个;代换片段长度最小为0.55 cM,最大为33.00 cM,平均长度为12.36 cM,总覆盖长度为247.20 cM,覆盖率为16.21%。利用BC2F2群体对14个产量相关性状进行QTL定位,共检测到控制8个性状的20个QTL。对性状表型值起增效作用的有11个,占总检出数的55%。控制产量相关性状的QTL存在簇状分布现象,这与表型相关分析结果相符合。