|Title:||分子標誌因子在蔬菜品種改良上之利用||Other Titles:||Utilization Of Molecular Markers In Vegetable Crop Improvement||Authors:||陳榮芳
Long-Fang O. Chen
|Keywords:||同功異構酶;限制內切酶酶譜多形性;聚合酶連鎖反應;逢機擴增多形性DNA;連鎖圖;數量性狀;Isozyme;Restriction fragment length polymorphism;Arbitrary-primer;Polymerase chain reaction;Random amplified polymorphic DNA;Quantitative trait;linkage map||Issue Date:||Jun-1993||Publisher:||農業試驗所||Related Publication(s):||農業試驗所特刊第41號||Start page/Pages:||103-120||Source:||蔬菜生產與發展研討會專刊||Conference:||蔬菜生產與發展研討會||Abstract:||
生物技術中應用於作物改良者，分子標誌因子（molecular markers）是較為實際與典型的例子之一。在蔬菜作物中，番茄分子標誌因子之研究堪稱典範。在可資利用之分子標誌因子來源中，隨著分子生物技術之進步，已由早期利用蛋白質、酵素膠體電泳法，配合酵素活性染色的同功異構酶（isozyme）分析，進而利用DNA限制內切酶酶譜多形性（Restriction fragment length polymorphisms；RFLPs)，至新近應用聚合酶連鎖反應（Polymerase chain reaction；PCR）所產生的聚合酶逢機擴增多形性DNA(Random amplified polymorphic DNA；RAPD）為標誌因子。這三種方法中，同功異構酶之發展最早，在蔬菜作物之報告亦不勝枚舉，其法之優點為分析方法簡單，而缺點則為可當分子標誌因子之因子座有限；而RFLP 分析法係利用DNA 限制內切酶辨識特殊鹽基系列之特性，若不同個體之DNA序列不同時，限制酶切割後所產生片段之大小、數目亦因此而有不同，經與該段序列有某種程度相似性之探針複合（hybridization ）檢定，所產生不同之圖譜做為標誌因子，此方法之優點為所產生之因子數遍佈整個基因組，缺點為分析所用之方法較耗人力與時間；目前蔬菜作物除番茄已建立良好之RFLP分子標誌圖譜外，馬鈴薯、萵苣、甜菜、豌豆及一些十字花科蕓苔屬類蔬菜亦已有了報告。而有關利用聚合醃連鎖反應，其逢機擴增DNA片段大小不同為標誌因子之方法，其優點為簡單、快速，缺點為大部分RAPD分子標誌因子為顯性，無由分辨異型合子，且有關RAPD在作物上利用之報告目前十分有限，在發展上是否有其他受限之處有待進一步之探討。此外分子標誌因子之利用亦已由早期用以篩選一些主效因子，如抗病、抗蟲、雄不稔性等因子之選拔，進一步用來標定數量性狀之選拔。
The utilization of molecular markers for crop improvement is believed to be one of the benefits from biotechnology. Presently, at least three types of molecular markers such as isozyme, RFLP (Restriction Fragment Length Polymorphism) and RAPD (Random Amplified ploymorphic DNA) derived from AP-PCR (Arbitrary Primer Polymerase Chain Reaction) are available. Among these, isozyme markers were studied the earliest and a lot of research has been conducted. The advantages of isozyme as molecular marker are its simplicity and ease for analysis. However, the limitation on the number of polymorphic loci is the major drawback in its application. RFLP markers analyzed on the DNA level are believed to be very promising because the markers obtained can be saturated all over the linkage map. Nevertheless, the profoundity and complexity of its methodologies have slowed down its utilization. Among the vegetable crop species, RFLP markers and linkage map have been well established in tomato. Other crops such as potato, lettuce, sugar beet, pea and Brassica species also have several studies on the RFLP markers. RAPD derived from AP-PCR was most recently reported. The advantages of RAPD markers are its rapidity and efficiency during the analyzing processes. However, the predominance with dominant gene action on RAPD markers will require twice as many markers as those of codominant markers to distinguish a heterozygote. Since only very limited studies are avaliable now on RAPD in crop species, whether there is any other limitation requires further investigation. Furthermore, gene mapping is no longer limited to those of major qualitative genes such as disease, insect resistance, and male sterility, etc. Selection for quantitative traits have become possible through the aid of molecular markers.
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