粳稻与几个广亲和品种杂交F1的花药培养效果

对粳稻品种秋光与几个广亲和品种及对照品种秋光的杂种F1进行花药培养,并对其中的一个组合所产生的白化苗做了PCR分析。结果表明：（1）秋光是一个培养力很高的品种,以秋光作为母本的粳／籼交组合的花药培养可以得到较高的培养力（绿苗率与愈伤组织诱导率的乘积）;（2）在供试的杂交组合中,培养力高低的一般趋势是：粳（籼）广亲和／籼（粳）广亲和＞粳／广亲和＞粳＞粳＞粳＞粳／籼;（3）与普通籼粳交相比,由于广亲和品种的参与提高了杂种F1的绿苗分化率, 白苗率;出愈率、分化率及培养力与出愈速度并无直接关系,选择适合的培养基并及时将愈伤组织转移到分化培养基上进行分化是提高花药培养的分化率和培养力的重要保证;（4）混生（或嵌合体）白化苗的形成是在分化初期愈伤组织发生遗传变异的结果,这种变异一经发生,无法用再分化的方法诱导绿苗的形成。     

影响水稻籼粳亚种间杂交构建回交自交系的花药培养效果的2个因素

水稻籼粳亚种间杂交构建的回交自交系（BIL）的10个株系在3种诱导培养基、1种分化培养基以及样株剪叶与否处理的结果表明,通用培养基（GM）不适合于籼粳交后代BIL的花药培养。N6和SK3都有较高的出愈率,但N6的绿苗发生率比SK3高,说明N6的愈伤组织质量和诱导效果更优于SK3。无论N6还是SK3,剪去样株叶片的绿苗发生率显著下降,显示花药培养过程中绿叶的作用很重要。完善籼粳亚种间杂交构建BIL的花药培养体系和从较多的株系中取材可保证花药培养的后代有较大的二倍体群体以供选择。     

籼粳分类的形态指数及其相关鉴定性状的遗传分析

利用通过典型籼粳交F1代加倍所构建的一个DH群体为材料,考察了121个DH株系的程氏指数及其相关的6个鉴定性状,并进行了数量性状座位(QTLs)分析.分别检测到叶毛的2个主效基因、谷粒长宽比的3个QTLs、稃色的1个QTL、稃毛的1个QTL、穗轴1～2节长的2个QTLs及酚反应的1个主效基因和2个QTLs.另外检测到与形态指数有关的4个QTLs,分别在第1,3,4和6染色体上.其中第1,3和6染色体上控制形态指数的3个QTLs分别与鉴定性状的部分QTLs位于同一区间.     

籼稻花药培养的研究——提高愈伤组织诱导率与分化率及根芽分化

3年来通过对籼稻(Oryza sativa subsp.hsien)花药培养的研究,愈伤组织的诱导率从0.2%提高到平均为2.85%,最高达18.75%;绿苗分化率从5%以下提高到平均为13.1%最高达到45.5%。在6个籼稻品种、38个籼×籼杂种中成功地诱导出了绿苗。本文着重研究了提高诱导率的有关因素。以单核晚期花粉的花药愈伤组织诱导频率最高。籼稻花粉去分化过程相对地要求较低浓度的培养基,籼稻花药对高浓度的改良 RM-1964培养基完全没反应。用杂种的诱导率较亲本材料为高。以易于诱导愈伤组织或易于分化绿苗的品种作亲本的杂种后代,诱导率与绿苗分化率亦较高。籼稻形成愈伤组织的高峰是在接种后的50—60天之间。愈伤组织分化的高潮是在转移后10—30天之间。籼稻愈伤组织的分化以先分化根的类型最多,在先分化根的愈伤组织中有36.74%能再分化出芽来。     

不同培养基对水稻胚性愈伤组织诱导及分化的影响

以水稻成熟胚为材料诱导愈伤组织,统计在不同基本培养基上的愈伤诱导率以及绿苗分化率,分析不同基本培养基及外源激素的含量和比例对愈伤组织生长及分化的影响。结果表明,试验材料对基本培养基具有选择性,MS培养基对籼稻种胚愈伤的诱导培养效果较好,NB培养基则更适合粳稻种胚愈伤的诱导培养;诱导继代培养基中加入多种氨基酸组合可有效提高出愈率和分化率,特别是粳稻的愈伤组织的诱导和分化需要多种氨基酸的共同作用;不同基因型水稻材料对激素和氨基酸组合的需求不同。     

利用RFLP标记定位水稻重要农艺性状的主效基因与微效基因

以一对籼粳交(圭630/02 428)经花药培养产生的双单倍体(DH)群体构建了含233个RFLP标记、覆盖基因组约2070cM的水稻分子图谱.利用QTL区间作图对水稻重要农艺性状如抽穗期、株高、有效穗、着粒数、实粒数、结实率和千粒重等7个性状的基因座位进行了定位分析,检测到8个主效基因和29个微效基因.双亲间性状差异较大有利于主效基因的鉴别.     

籼稻花药培养研究——Ⅰ.基本培养基及附加成分在诱导籼稻花药产生愈伤组织及根芽分化中的作用

对6个籼稻(oryza sativa Subsp.Shien)品种和39个籼×籼杂种的花药在离体条件下进行培养,有5个品种及35个杂种得到了愈伤组织,平均诱导率为2.18％。在3个品种及11个籼×籼杂种中得到了绿苗或绿芽。本文着重报道基本培养基及其附加成分在诱导籼稻花药产生愈伤组织及根芽分化中的作用。 1.试验了几种诱导愈伤组织的培养基,以Miller培养基 2,4—D2毫克/升 酵母浸出液1,000毫克/升 激动素1毫克/升 吲哚乙酸2毫克/升 椰乳15％为最好。诱导率高者可达11—15％,平均诱导率在3％以上。 2.Ms、Nitsch及Miller培养基均可诱导籼稻花药愈伤组织分化出绿色的花粉植株。 3.籼稻花药愈伤组织的分化,随着激动素/生长素比值的增高,绿苗分化率及总分化率均有提高的趋势。而粳稻的这种关系不甚明显。 4.Miller培养基附加2毫克/升的吲哚乙酸对促进具茎、叶而无根的籼稻花粉小植株产生根有很好的作用。在这种培养基上,不仅可以诱导根的发生,而且根系发达,生长较弱的苗转移到这种培养基后,因根系健壮,生势好,转入盆栽,基本可以全部成活。     

影响水稻花药培养力的数量性状基因座位间的互作

控制数量性状的多个基因间不仅存在加性效应,还存在非等位基因间的互作。对一个籼粳交后代的加倍单倍体群体花药培养,通过Ｅｐｉｓｔａｔ软件分析影响水稻花药 数量性状基因座位间的互作。结果表明,愈伤组织诱导率主要受两个单基因的影响,不存在双基因条件型互作,但有２对互适型互作与其有关。     

粳糯杂交组合花培植株的糯性遗传及育种效果

水稻花培的绿苗成功率是权衡花药培养效率及其应用价值的重要指标之一。 研究表明,不同稻种类型配组的绿苗成功率的顺序是:粳/糯>粳/粳>粳/籼>A/R(籼型杂交稻)>籼/籼。我们利用粳/糯绿苗成功率较高的特性,从3个粳糯杂交组合的花药培养中,诱导出230多株绿苗,通过鉴定选出4个花培糯稻品系。     

用RFLP标记剖析水稻穗颈维管束及穗部性状的遗传基础

采用籼、粳亚种间杂种F1(圭630×02428)花药培养获得的DH群体,对水稻穗颈大、小维管束数和倒数第2节间大、小维管束数等4个维管束性状,以及一、二次枝梗数,每穗颖花数3个穗部性状进行QTL分析,共检测到16个QTLs,其中有7个QTLs的加性效应较大,单个QTL的贡献率在20%以上。发现有4个QTLs成簇分布于第1染色体从RZ776到C11的大约35cM的区段上,来源于亲本"圭630"的这一染色体区段对穗颈大维管束、第2节间大维管束、第2节间小维管束和二次枝梗数4个性状的表达均具有增效作用。还讨论了利用分子标记辅助选择聚合增效QTLs、实现穗颈维管束性状遗传改良的策略。     

Analysis of quantitative trait loci which contribute to anther culturability in rice (Oryza sativa L.)

Anther culturability of rice is significantly different between indica and japonica varieties. A doubled haploid (DH) population was established via anther culture of an indica/japonica hybrid on SK₃ medium, which had been shown particularly suitable for anther culture of indica/japonica hybrids. For analyzing the quantitative trait loci (QTLs) responsible for anther culturability, anthers of the DH lines were again cultured with SK₃ medium and parameters for four traits representing the anther culturability were surveyed and analyzed with the molecular map constructed from the same DH population. The parameters for four major traits were as follows: callus induction frequency (CI), green plantlet differentiation frequency (GPD), albino plantlet differentiation frequency (APD), and green plantlet yield frequency (GPY). All four traits displayed continuous distributions among the DH lines. The correlation coefficients between these traits were also tested and showed that there was no relationship between callus induction and green plantlet differentiation frequencies, but both showed strong positive correlation with the frequency of green plantlet yield. For callus induction frequency, five QTLs were identified on chromosomes 6, 7, 8, 10 and 12. Two QTLs for green plantlet differentiation frequency were located on chromosomes 1 and 9. There was a major QTL for albino plantlet differentiation frequency on chromosome 9. No independent QTL was found for green plantlet yield frequency. The results may be useful in the selection of parents with high response to anther culture for rice haploid breeding and in the establishment of permanent DH populations for molecular mapping.     

用花药培养创建小麦加倍单倍体作图群体

用花药培养创建加倍单倍体作图群体是构建数量性状基因作图群体的有效途径,但通过花药培养生产小麦加倍单倍体的成功率较低。该文考查了小麦幼穗发育时期、低温处理幼穗及高温处理接种花药对花粉愈伤诱导率的影响。采用春播小麦做为花药供体,推迟花药接种时间,避免试管苗在低温条件下越夏,有利于早移栽,培育冬前壮苗,同时加强试管苗移栽后的田间管理,保证安全越冬,有效地提高了花药培养生产加倍单倍体的成功率。通过花药培养创建了一个具有１９１个个体的小麦加倍单倍体作图群体,该群体将用于小麦有关抗旱性状及产量性状基因的遗传作图     

水稻籼粳杂种生殖障碍的基因定位分析

籼稻(Oryza sativa L.ssp.indica)与粳稻(O.sativa ssp.japonica)杂交优势明显但存在生殖隔离。生殖障碍主要表现为胚囊败育、花粉败育、开花时花药不开裂和雌雄异熟。应用具有137个标记位点的籼、粳杂交(“窄叶青8号”/“京系17”)F_1花药培养获得的127个双单倍体(DH)群体构建的RFLP图谱,对控制籼、粳杂种小穗败育的基因座位进行了定位研究。结果在第1、3、4、5、6、7、8、12染色体上检测到10个基因座位,其中第3、12染色体上的2个不育基因位点stj-3和stj-12与同一杂交组合F_2分离群体中发现的异常分离热点处于相同的染色体区段。Ssj-6的基因加性效应为负值,有增加籼、粳亲和性的作用;其余的不育基因座位皆有增加籼、粳杂种不育性的作用。     

Antimitotic and hormone effects on green double haploid plant production through anther culture of Mediterranean japonica rice

Rice double haploid (DH) plants are produced mainly through anther culture. In order to improve the anther culture protocol, microspores of two japonica rice genotypes (NRVC980385 and H28) were subjected to three growth regulator combinations and four colchicine treatments on induction medium. In addition, a post anther culture procedure using colchicine or oryzalin was tested to induce double haploid plantlets from haploid plantlets. A cold pre-treatment of microspores for 9 days at 10 °C increased callus induction 50-fold in the NRCV980385 genotype. For both genotypes, 2 mg L^?1 2,4-D and 1 mg L^?1 kinetin on colchicine-free induction medium gave the best culture responses. The culturability of both genotypes changed on colchicine-supplemented induction media. A high genotype dependency was recorded for callus induction, callus regenerating green plantlets and regeneration of green double haploid plantlets. Colchicine at 300 mg L^?1 for 48 h enhanced callus induction 100-fold in H28. Colchicine-supplemented media clearly improved green double haploid plantlet regeneration. We showed that the post-anther culture treatment of haploid plantlets at 500 mg L^?1 of colchicine permitted fertile double haploid plantlets to be generated. Finally, an enhanced medium-throughput flow cytometry protocol for rice was tested to analyse all the plantlets from anther and post anther culture.     

Genetic analysis of morphological index and its related taxonomic traits for classification of indica/japonica rice

A doubled haploid population derived from anther culture of ZYQ8/JX17 F1, a typical indica and japonica hybrid, was used in this study. Morphological index and its related taxonomic traits were investigated in 121 DH lines. The quantitative trait loci (QTLs) for morphological index and its related taxonomic traits were analyzed. Two major QTLs for leaf hairiness, three QTLs for length/width of grain, one QTL for color of hull when heading, one QTL for hairiness of hull, two QTLs for length of the first and second panicle internode, and one major QTL and two QTLs for phenol reaction were detected. Four QTLs for morphological index were also identified on chromosomes 1, 3, 4 and 6, respectively, three of which on chromosomes 1, 3 and 6, respectively, were found to be located in the same chromosome regions where some QTLs for the related taxonomic traits were located.     

Genetic analysis of morphological index and its related taxonomic traits for classification of indica/japonica rice

A doubled haploid population derived from anther culture of ZYQ8/JX17 F₁, a typical indica and japonica hybrid, was used in this study. Morphological index and its related taxonomic traits were investigated in 121 DH lines. The quantitative trait loci (QTLs) for morphological index and its related taxonomic traits were analyzed. Two major QTLs for leaf hairiness, three QTLs for length/width of grain, one QTL for color of hull when heading, one QTL for hairiness of hull, two QTLs for length of the first and second panicle internode, and one major QTL and two QTLs for phenol reaction were detected. Four QTLs for morphological index were also identified on chromosomes 1, 3, 4 and 6, respectively, three of which on chromosomes 1, 3 and 6, respectively, were found to be located in the same chromosome regions where some QTLs for the related taxonomic traits were located.     

Genetic analysis of leaffolder resistance in rice

A double haploid(DH)population,which consists of 120 lines derived from anther culture of a typical indica and japonica hybrid'CJ06'/'TN1',was used to investigate the genetic basis for rice leaffolder resistance.Using a constructed molecular linkage map,five QTLs for rolled leaves were detected on chromosomes 1,2,3,4,and 8.The positive alleles from C J06 on chromosomes 3,4,and 8 in-creased the resistance to dee leaffolder,and the alleles from TN1 on chromosomes 1 and 2 also enhanced resistance to leaffolder.The interactions between QTLs were identified and tested,and four conditional interactions were acquired for resistance to rice leaffolder.These loci were located on chromosomes 2,9,10,and 11,respectively.QTL pyramiding indicated that the positive alleles affect resis-tance to leaffolder.The prospective application of this data in rice breeding was also discussed.     

甲基磺酸乙酯对水稻花药离体培养效应的研究

本文以粳稻“105”、籼稻“华03”和“鄂早6号”为实验材料,研究了不同浓度化学诱变剂EMS处理对离体培养水稻花药中花粉细胞脱分化与再分化和培养初期花药呼吸作用的影响。结果表明,低浓度EMS能显著提高花药愈伤组织诱导率,高浓度EMS则有明显的抑制作用。EMS对培养初期花药呼吸作用的影响与花药愈伤组织诱导率之间存在明显的平行关系。     

Quantitative trait loci mapping associated with plant regeneration ability from seed derived calli in rice (Oryza sativa L.)

Quantitative trait loci (QTLs), which are associated with the ability of plant regeneration from seed derived calli, were detected using a recombinant inbred (RI) population from a cross between 'Milyang 23 (toingil)' and 'Gihobyeo (japonica)' in rice (Oryza sativa L.). A tongil type cultivar, 'Milyang 23', has a lower frequency of callus induction and plant regeneration than those of japonica 'Gihobyeo'. Transgressive segregations were observed for the callus induction rate and plant regeneration ability from seed derived calli of the RI population. An interval mapping analysis was used to identify the QTL controlling the plant regeneration ability. Two QTLs for the callus induction rate were detected on chromosomes 1 and 2, explaining the 10.9% total phenotypic variation. Four QTLs that are associated with the plant regeneration ability were located on chromosomes 2, 3, and 11, accounting for 25.7% of the total phenotypic variation.