权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Utility of minor somaclonal variation and detection of the genetic mutation in rice.

水稻微小体克隆变异的实用性和基因突变的检测。

基本信息

批准号：
10660005
负责人：
ABE Toshinori
金额：
$ 0.7万
依托单位：
YAMAGATA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10660005/
关键词：
Oryza sativa L.Somaclonal variation short culm Chloroplast DNA Southern hybridization DNA Delation Oryza sastiva L.AFLP 培養変異 PCR フィンガープリント欠失

项目摘要

1. Somaclomnal variation obtained from short term culture of rice calli. Somaclonal variation obtained from tissue culture regenerants in expected for crop improvement. We obtained 10 regenerated plants from seed calli of japonica variety, Reimai. The plants were grown in green house until mature stage and analyzed agronomic traits. Out of 10 regenerants, 3 plants were 8-10 cm shorter that Reimei and other regenerated plants. While, one regenerated plant showed fewer ripened grains and lower ripening percentage. The other characters were almost the same. From these results, It should be clear the genes controlling the culm length in these regenerated plants.2. Detection of genomic change at molecular level coused by long term culture of rice calli. Chloroplast DNA (ctDNA) alteration of long term callus cultures derived from rice seeds was easily detected when rice chloroplast DNA fragment BamHI-1 was used as a probe. Chloroplast DNA (ctDNA) molecules of two callus lines out of 12 callus lines cultured for 36 months were deficient in large regions of the rice chloroplast genome. Maps of deleted ctDNA molecules were constructed using a seventeen hybridization probes. In the callus lines of T36-1 and T36-2, small sizes of ctDNA molecules (58.5 and 7.5 kb) were mapped. When these two callus lines were further cultured on the MS maintenance medium for more six months, further deletion (about 0.5 kb) was occurred in one callus line (T42-2). From the restriction maps of the deleted derivatives of ctDNA, callus lines of T36-1 and T42-1 lost both large inverted repeat sequences, together with the intervening small single copy region. On the other hand, callus lines of T36-2 and T42-2 lost rbcL and most of the large single copy region. These ctDNA molecules commonly retain only about 20 kb that encode tm gene cluster and rpo genes. These results suggests that this region of rice chloroplast genome contains at least one functional origin of replication.

1.从水稻愈伤组织的短期培养中获得的体细胞变异。从组织培养再生体中获得的体细胞克隆变异有望改善作物。我们从粳稻品种灵迈的种子愈伤组织中获得了10株再生植株。将植物在温室中生长直至成熟阶段并分析农艺性状。 10株再生植物中，有3株比Reimei和其他再生植物矮8-10厘米。然而，一株再生植株的成熟粒数较少，成熟率较低。其他角色几乎是一样的。从这些结果应该可以清楚控制这些再生植株的秆长度的基因。 2．检测水稻愈伤组织长期培养引起的分子水平基因组变化。当使用水稻叶绿体 DNA 片段 BamHI-1 作为探针时，可以轻松检测到源自水稻种子的长期愈伤组织培养物的叶绿体 DNA (ctDNA) 变化。 Chloroplast DNA (ctDNA) molecules of two callus lines out of 12 callus lines cultured for 36 months were deficient in large regions of the rice chloroplast genome.使用 17 个杂交探针构建了删除的 ctDNA 分子图谱。在 T36-1 和 T36-2 的愈伤组织系中，绘制了小尺寸的 ctDNA 分子（58.5 和 7.5 kb）。当这两个愈伤组织系在MS维持培养基上进一步培养六个月以上时，一个愈伤组织系(T42-2)中发生进一步缺失(约0.5kb)。从删除的 ctDNA 衍生物的限制性图谱来看，T36-1 和 T42-1 的愈伤组织系丢失了两个大的反向重复序列，以及中间的小单拷贝区域。另一方面，T36-2和T42-2的愈伤组织系丢失了rbcL和大部分大的单拷贝区域。这些 ctDNA 分子通常仅保留约 20 kb 的长度，用于编码 tm 基因簇和 rpo 基因。这些结果表明，水稻叶绿体基因组的这一区域包含至少一个功能性复制起点。