TILLING for allergen reduction and improvement of quality traits in peanut (Arachis hypogaea L.)

Knoll, Joseph E.; Ramos, M. Laura; Zeng, Yajuan; Holbrook, C. Corley; Chow, Marjorie; Chen, Sixue; Maleki, Soheila; Bhattacharya, Anjanabha; Ozias-Akins, Peggy
January 2011
BMC Plant Biology;2011, Vol. 11 Issue 1, p81
Academic Journal
Background: Allergic reactions to peanuts (Arachis hypogaea L.) can cause severe symptoms and in some cases can be fatal, but avoidance is difficult due to the prevalence of peanut-derived products in processed foods. One strategy of reducing the allergenicity of peanuts is to alter or eliminate the allergenic proteins through mutagenesis. Other seed quality traits could be improved by altering biosynthetic enzyme activities. Targeting Induced Local Lesions in Genomes (TILLING), a reverse-genetics approach, was used to identify mutations affecting seed traits in peanut. Results: Two similar copies of a major allergen gene, Ara h 1, have been identified in tetraploid peanut, one in each subgenome. The same situation has been shown for major allergen Ara h 2. Due to the challenge of discriminating between homeologous genes in allotetraploid peanut, nested PCR was employed, in which both gene copies were amplified using unlabeled primers. This was followed by a second PCR using gene-specific labeled primers, heteroduplex formation, CEL1 nuclease digestion, and electrophoretic detection of labeled fragments. Using ethyl methanesulfonate (EMS) as a mutagen, a mutation frequency of 1 SNP/967 kb (3,420 M2 individuals screened) was observed. The most significant mutations identified were a disrupted start codon in Ara h 2.02 and a premature stop codon in Ara h 1.02. Homozygous individuals were recovered in succeeding generations for each of these mutations, and elimination of Ara h 2.02 protein was confirmed. Several Ara h 1 protein isoforms were eliminated or reduced according to 2D gel analyses. TILLING also was used to identify mutations in fatty acid desaturase AhFAD2 (also present in two copies), a gene which controls the ratio of oleic to linoleic acid in the seed. A frameshift mutation was identified, resulting in truncation and inactivation of AhFAD2B protein. A mutation in AhFAD2A was predicted to restore function to the normally inactive enzyme. Conclusions: This work represents the first steps toward the goal of creating a peanut cultivar with reduced allergenicity. TILLING in peanut can be extended to virtually any gene, and could be used to modify other traits such as nutritional properties of the seed, as shown in this study.


Related Articles

  • Genome Sequencing and Analysis of the Peanut B-Genome Progenitor (Arachis ipaensis). Lu, Qing; Li, Haifen; Hong, Yanbin; Zhang, Guoqiang; Wen, Shijie; Li, Xingyu; Zhou, Guiyuan; Li, Shaoxiong; Liu, Hao; Liu, Haiyan; Liu, Zhongjian; Varshney, Rajeev K.; Chen, Xiaoping; Liang, Xuanqiang // Frontiers in Plant Science;5/3/2018, p1 

    Peanut (Arachis hypogaea L.), an important leguminous crop, is widely cultivated in tropical and subtropical regions. Peanut is an allotetraploid, having A and B subgenomes that maybe have originated in its diploid progenitors Arachis duranensis (A-genome) and Arachis ipaensis (B-genome),...

  • Peanut genome sequence offers significant benefits. Byrns, Steve // Southwest Farm Press Exclusive Insight;4/24/2014, p5 

    The article reports on the launch of a peanut genome sequence by the International Peanut Genome Initiative and its impact on the industry and consumers in 2014.

  • The polyembryo gene ( OsPE) in rice. Puri, Anju; Basha, P. Osman; Kumar, Mankesh; Rajpurohit, Deepak; Randhawa, Gursharn S.; Kianian, Shahryar F.; Rishi, Anantharama; Dhaliwal, Harcharan S. // Functional & Integrative Genomics;Aug2010, Vol. 10 Issue 3, p359 

    T-DNA insertional mutagenesis is one of the most important approaches for gene discovery and cloning. A fertile polyembryo mutant generated by T-DNA/ Ds insertion in Oryza sativa, cv. Basmati 370 showed twin or triple seedlings at a frequency of 15–20%. T-DNA insertion was confirmed by...

  • Genetic diversity of peanut (Arachis hypogaea L.) and its wild relatives based on the analysis of hypervariable regions of the genome. De Carvalho Moretzsohn, Marcio; Hopkins, Mark S.; Mitchell, Sharon E.; Kresovich, Stephen; Montenegro Valls, Jose Francisco; Ferreira, Marcio Elias // BMC Plant Biology;2004, Vol. 4, p1 

    Background: The genus Arachis is native to a region that includes Central Brazil and neighboring countries. Little is known about the genetic variability of the Brazilian cultivated peanut (Arachis hypogaea, genome AABB) germplasm collection at the DNA level. The understanding of the genetic...

  • The peanut genome -- making a better peanut. Robinson, Elton // Delta Farm Press Exclusive Insight;7/ 8/2014, p1 

    The article reports on sequencing of the peanut genome by the International Peanut Genome Initiative, a multinational group of crop geneticists which has worked in cooperation to accomplish the task as of 2014. The topics discussed include the collection of wild species of peanuts including...

  • Impact of GMO'S on environment and human health. Godheja, Jai // Recent Research in Science & Technology;May2013, Vol. 5 Issue 5, p26 

    Genetic pollution is the term of genetics in which the genetic information is transferred in to the organisms where it is not needed or where this information never existed before. Genetic pollution is a controversial[1][2] term for uncontrolled[3][4] gene flow into wild populations. This flow...

  • Reverse genetics techniques: engineering loss and gain of gene function in plants. Gilchrist, Erin; Haughn, George // Briefings in Functional Genomics;Mar2010, Vol. 9 Issue 2, p103 

    Genetic analysis represents a powerful tool that establishes a direct link between the biochemical function of a gene product and its role in vivo. Genome sequencing projects have identified large numbers of plant genes for which no role has yet been defined. To address this problem a number of...

  • Mutant Resources for the Functional Analysis of the Rice Genome. Wang, Nili; Long, Tuan; Yao, Wen; Xiong, Lizhong; Zhang, Qifa; Wu, Changyin // Molecular Plant (Oxford University Press / USA);May2013, Vol. 6 Issue 3, p596 

    This review describes in detail the present status of mutant resource characteristics, availability for rice mutant resources and tagged gene database. It can be an important reference for rice functional genomics research.Rice is one of the most important crops worldwide, both as a staple food...

  • CRISPR/Cas9-Mediated Targeted Mutagenesis in the Liverwort Marchantia polymorpha L. Sugano, Shigeo S.; Shirakawa, Makoto; Takagi, Junpei; Matsuda, Yoriko; Shimada, Tomoo; Hara-Nishimura, Ikuko; Kohchi, Takayuki // Plant & Cell Physiology;Mar2014, Vol. 55 Issue 3, p475 

    Targeted genome modification technologies are key tools for functional genomics. The clustered regularly interspaced short palindromic repeats (CRISPR)-associated endonuclease Cas9 system (CRISPR/Cas9) is an emerging technology for targeted genome modification. The CRISPR/Cas9 system consists of...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics