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研究生(外文):Widyatani Anggriana
論文名稱(外文):Self-incompatibility study in petunia
指導教授(外文):Yueh-Long Chang
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自交不親和性是許多開花植物用來防止近親繁殖的方式,並且促進雜交授粉.所有的配子型和孢子型自交不親和性系統可以有效地防止自交.在矮牽牛中,自交不親和性類屬於茄科自交不親和性繁殖障礙因子,是由眾多多型性的S等位基因所調控.雌性決定S基因轉譯S-RNA酶.S-RNA酶包含所需的花粉排斥反應和參與等位基因特異性識別高變異區的RNA酶的活動區域. 在本研究中,DNA是從三個矮牽牛商業雜交系中萃取。DNA指紋圖譜是藉由ISSR技術來建立。ISSR的引子是從微結構的序列的特徵設計而來,用於擴增與引子相對應的基因組區域。這些標記可用於直接測量DNA的遺傳多樣性,可區分三個自交不親和的商業品系花粉混合產生的子代。DNA特徵的結果可以提供在自交不親和植物中,促使自交系可經由花粉混合策略應用而產生。 克服自交不親和性(SI)的屏障,可以藉由三個雄性花粉混合法產生自花授粉的後代。
Self Incompatibility (SI) is a reproductive strategy adopted by many flowering plant species to prevent inbreeding and promote cross-pollination. All the gametophytic and sporophytic self-incompatibility systems are therefore efficient in preventing selfing. In petunia, its self-incompatibility system includes Solanaceae type SI in which this reproductive barrier is regulated by highly polymorphic S alleles. The female-determinant S genes encode S-RNAse. S-RNAse contains an RNAse activity domain required for pollen rejection and a hypervariable region involved in allele specific recognition. In this study, genomic DNA was extracted from three petunia hybrid lines. DNA fingerprinting was performed by the inter simple sequence repeat analysis (ISSR). ISSR primers are designed from sequences of microsatellite structures and used for amplifying genomic regions that are flanked by primers. These markers can be used to directly measure genetic diversity at the DNA level for progeny individuals derived from the self-incompatible plant pollinated with a mixture of pollens collected from three commercial hybrid lines. The results of DNA fingerprinting can provide the evidence to make sure that self-pollination can occur in self-incompatible plants using pollen mixture strategy. Self-incompatibility (SI) barrier might be overcomed by using pollen mixture of three male donors for producing self-pollinated progeny
Abstract i
Acknowledgement iii
List of content iv
List of figure vi
I. Introduction 1
II. Material and method
2.1 Material 10
2.2 Method 10
2.2.1 Pollination 10
2.2.2 Pollen germination 11
2.2.3 DNA isolation
A. DNA isolation and extraction 11
B. DNA purification 12
C. Quantification and qualification of genomic DNA 13
2.2.4 Analysis ISSR 14
2.2.5 Aniline blue staining of pollen tubes within style 14
III. Result
3.1 Self-Incompatibility (SI) status of 13 plants 15
3.2 Pollen germination rate test 15
3.3 Pollen tube growth 17
3.4 Analysis ISSR 18
3.5 Genetic diversity 18
IV. Discussion 20
Figure 28
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