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Developing a Peanut (Arachis hypogae...

University of Florida.

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Developing a Peanut (Arachis hypogaea L.) Transformation Protocol Using Agrobacterium and Characterizing Late Leaf Spot Disease Resistance in Peanut.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Developing a Peanut (Arachis hypogaea L.) Transformation Protocol Using Agrobacterium and Characterizing Late Leaf Spot Disease Resistance in Peanut./
Author:	Hsieh, Yih-Feng.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2016,
Description:	154 p.
Notes:	Source: Dissertations Abstracts International, Volume: 79-03, Section: B.
Contained By:	Dissertations Abstracts International79-03B.
Subject:	Molecular biology. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10646380
ISBN:	9780355196320

Developing a Peanut (Arachis hypogaea L.) Transformation Protocol Using Agrobacterium and Characterizing Late Leaf Spot Disease Resistance in Peanut.
Hsieh, Yih-Feng.

Developing a Peanut (Arachis hypogaea L.) Transformation Protocol Using Agrobacterium and Characterizing Late Leaf Spot Disease Resistance in Peanut. - Ann Arbor : ProQuest Dissertations & Theses, 2016 - 154 p.

Source: Dissertations Abstracts International, Volume: 79-03, Section: B.

Thesis (Ph.D.)--University of Florida, 2016.

This item must not be sold to any third party vendors.

Efficient and genotype-independent in vitro regeneration is an essential prerequisite for incremental trait improvement in peanut ( Arachis hypogaea L.) via genetic transformation. We have optimized a facile and rapid method to obtain direct shoot organogenesis from cotyledonary node (CN) explants excised from peanut seedlings germinated on cytokinin-supplemented Murashige and Skoog (MS) basal salt medium. Starting with mature embryos, shoot induction occurred in approximately 7 weeks, followed by 4 weeks for rooting of excised shoots and 3 weeks of acclimatization of regenerated plantlets in soil. The regeneration and transformation system described here is time-efficient, yielding greenhouseacclimatized plantlets within 14 weeks, in contrast to 12-14 months required for initiating and regenerating somatic embryogenic cultures, currently the most tractable method available for peanut transformation. The highest shoot induction frequency and shoot quality was obtained with 6.66 µM 6-Benzylaminopurine, followed by adequate root induction with 5.37 µM α-Naphthaleneacetic acid. New Mexican Valencia A was chosen for Agrobacterium-mediated transformation. The transformation frequency were 1.25 % and 2.43% with pWBvec10a and pSag12::IPT, respectively. The CN-based direct regeneration system described here is time-efficient and amenable to Agrobacterium-mediated transformation, and therefore should be further explored for peanut transgenic improvement. Late leaf spot (LLS) disease is one of the most severe diseases of peanut worldwide. However, the molecular network related to disease resistance is still unknown. This study provided evidence that associated disease development with signaling networks in response to LLS using microscopic observation and semi-reverse transcription polymerase chain reaction (Semi-RT-PCR). The results showed that at least 15 days were required for symptom development on peanut leaves after inoculation. LLS spore germination was inhibited at the early infection stage (0 to 3 DAI) and sporulation was low in the resistant cultivar. Furthermore, the genes involved in hormone signaling transduction and defense responses after LLS infection that were analyzed through Semi RT-PCR showed significant differences in expression between resistant and susceptible peanut cultivars, especially the resveratrol synthase gene, indicating that phytoalexin biosynthesis might play an important role in LLS resistance. Differentially expressed genes could be further applied as molecular markers in peanut breeding programs for selecting LLS disease resistant cultivars.

ISBN: 9780355196320Subjects--Topical Terms:

517296
Molecular biology.
Subjects--Index Terms:

Agrobacterium-mediated transformation

Developing a Peanut (Arachis hypogaea L.) Transformation Protocol Using Agrobacterium and Characterizing Late Leaf Spot Disease Resistance in Peanut.
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300 $a 154 p.
500 $a Source: Dissertations Abstracts International, Volume: 79-03, Section: B.
500 $a Publisher info.: Dissertation/Thesis.
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506 $a This item must not be sold to any third party vendors.
520 $a Efficient and genotype-independent in vitro regeneration is an essential prerequisite for incremental trait improvement in peanut ( Arachis hypogaea L.) via genetic transformation. We have optimized a facile and rapid method to obtain direct shoot organogenesis from cotyledonary node (CN) explants excised from peanut seedlings germinated on cytokinin-supplemented Murashige and Skoog (MS) basal salt medium. Starting with mature embryos, shoot induction occurred in approximately 7 weeks, followed by 4 weeks for rooting of excised shoots and 3 weeks of acclimatization of regenerated plantlets in soil. The regeneration and transformation system described here is time-efficient, yielding greenhouseacclimatized plantlets within 14 weeks, in contrast to 12-14 months required for initiating and regenerating somatic embryogenic cultures, currently the most tractable method available for peanut transformation. The highest shoot induction frequency and shoot quality was obtained with 6.66 µM 6-Benzylaminopurine, followed by adequate root induction with 5.37 µM α-Naphthaleneacetic acid. New Mexican Valencia A was chosen for Agrobacterium-mediated transformation. The transformation frequency were 1.25 % and 2.43% with pWBvec10a and pSag12::IPT, respectively. The CN-based direct regeneration system described here is time-efficient and amenable to Agrobacterium-mediated transformation, and therefore should be further explored for peanut transgenic improvement. Late leaf spot (LLS) disease is one of the most severe diseases of peanut worldwide. However, the molecular network related to disease resistance is still unknown. This study provided evidence that associated disease development with signaling networks in response to LLS using microscopic observation and semi-reverse transcription polymerase chain reaction (Semi-RT-PCR). The results showed that at least 15 days were required for symptom development on peanut leaves after inoculation. LLS spore germination was inhibited at the early infection stage (0 to 3 DAI) and sporulation was low in the resistant cultivar. Furthermore, the genes involved in hormone signaling transduction and defense responses after LLS infection that were analyzed through Semi RT-PCR showed significant differences in expression between resistant and susceptible peanut cultivars, especially the resveratrol synthase gene, indicating that phytoalexin biosynthesis might play an important role in LLS resistance. Differentially expressed genes could be further applied as molecular markers in peanut breeding programs for selecting LLS disease resistant cultivars.
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