Transcriptome Comparison Reveals Distinct Selection Patterns in Domesticated and Wild Agave Species, the Important CAM Plants

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Transcriptome Comparison Reveals Distinct Selection Patterns in Domesticated and Wild Agave Species, the Important CAM Plants Hindawi International Journal of Genomics Volume 2018, Article ID 5716518, 12 pages https://doi.org/10.1155/2018/5716518 Research Article Transcriptome Comparison Reveals Distinct Selection Patterns in Domesticated and Wild Agave Species, the Important CAM Plants 1 2 1 3 1 1 Xing Huang , Bo Wang, Jingen Xi, Yajie Zhang, Chunping He, Jinlong Zheng, 4 4 4 1 1 Jianming Gao, Helong Chen, Shiqing Zhang, Weihuai Wu, Yanqiong Liang, 1 and Kexian Yi 1Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China 2National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, Hubei 430070, China 3Hainan Climate Center, Haikou 570203, China 4Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China Correspondence should be addressed to Xing Huang; [email protected] and Kexian Yi; [email protected] Received 27 February 2018; Revised 21 August 2018; Accepted 24 September 2018; Published 22 November 2018 Academic Editor: Giandomenico Corrado Copyright © 2018 Xing Huang et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Agave species are an important family of crassulacean acid metabolism (CAM) plants with remarkable tolerance to heat and drought stresses (Agave deserti) in arid regions and multiple agricultural applications, such as spirit (Agave tequilana) and fiber (Agave sisalana) production. The agave genomes are commonly too large to sequence, which has significantly restricted our understanding to the molecular basis of stress tolerance and economic traits in agaves. In this study, we collected three transcriptome databases for comparison to reveal the phylogenic relationships and evolution patterns of the three agave species. The results indicated the close but distinctly domesticated relations between A. tequilana and A. sisalana. Natural abiotic and biotic selections are very important factors that have contributed to distinct economic traits in agave domestication together with artificial selection. Besides, a series of candidate unigenes regulating fructan, fiber, and stress response-related traits were identified in A. tequilana, A. sisalana, and A. deserti, respectively. This study represents the first transcriptome comparison within domesticated and wild agaves, which would serve as a guidance for further studies on agave evolution, environmental adaptation, and improvement of economically important traits. 1. Introduction for fiber production in tropical regions [6]. As a native wild plant in the Sonoran Desert regions of the Southwestern Agave species assembled an important group of crassulacean United States and Northwestern Mexico, Agave deserti has acid metabolism (CAM) plants with remarkable tolerance to successfully survived in a severe environment within elevation heat and drought stresses in arid regions [1]. CAM plants ranges that experience both hot, dry summers and occasional usually have a higher efficiency in water use than C3 and freezing temperatures in winter [7, 8]. This capacity of high C4 plants [2]. For this reason, CAM plants brought a great tolerance to multiple stresses has important values to the chance to enhance sustainable production of food and bioe- improvement of the main food crops. These economic and nergy under the background of limited freshwater resources stress-tolerant features have made agave a model CAM crop and global climate change [3]. As a traditional cultivated system for a hot and dry/droughty/xeric environment [9]. CAM plant, Agave tequilana has been used for the produc- However, few reports have revealed the physiology and tion of distilled spirit tequila for centuries [4]. Further, it also molecular basis of agaves, especially in A. sisalana. To date, shows a great potential in bioenergy production [5]. Besides, A. tequilana-related studies mainly focused on fructan, with Agave sisalana has also been widely cultivated as a cash crop the obvious purpose of improving fructan production and 2 International Journal of Genomics 2 Agave americana 2 Agave terraccianoi 25 3 Agave deserti(former) 7 Agave sisalana Agave tequilana 2 Agave angustifolia 3 Agave macracantha Agave marmorata 7 Agave kerchovei 44 65 Agave stricta Agave potatorum 49 Agave karwinskii 3 Agave schidigera Agave subsimplex 40 Agave deserti(current) Agave cerulata Agave victoriae-reginae 0.0010 (a) 70 Agave sisalana Agave tequilana Agave americana Agave deserti 0.00020 (b) Figure 1: Phylogenetic analysis of chloroplast sequences in Agave species. (a) The trnL+trnL-trnF sequences (about 900 bp) were used to construct a phylogenetic tree. (b) The sequences of A. tequilana (GAHU01110124), A. deserti (GAHT01022741), and A. americana (KX519714) were from NCBI. A. sisalana (CL7065.Contig2) sequence and the other 13 sequences were from previous studies [15, 24]. Table 1: Summary of the transcriptomes for the 3 Agave species. Species A. tequilana A. deserti A. sisalana Total sequenced high-quality data 293.5 Gbp 184.7 Gbp 11.3 Gbp Number of Agave unigenes 204,530 128,869 131,422 Sum length of Agave unigenes 204.9 Mbp 125.0 Mbp 77.6 Mbp N50 length 1387 bp 1323 bp 861 bp application to generate bioenergy [5, 10, 11]. A. deserti- comparison has also been conducted for evolution analysis related studies were mainly conducted for its ecological and and searching economically important traits in some genome physiological adaptation to a severe environment, which is unavailable crops [22]. In this study, we selected three tran- highly valuable for the improvement of the main food crops scriptome databases for comparison to reveal the phylogeny [12]. Besides, a series of saponin-related researches have been and evolution pattern of the three agave species [23, 24]. reported, while few reports were related to fiber in A. sisalana Those genes related to species-specific traits would be also [13, 14]. These three agave species are closely related species identified and evaluated for their importance in agronomy but with different remarkable biological features. A previous production and environmental adaptation of agaves. This study has revealed the phylogenetic relations according to study represents the first transcriptome comparison within their trnL sequences [15]. The result showed that they were domesticatedandwildagaves,whichwouldserveasaguidance closely related species in spite of different origins for A. tequi- for further studies on agave evolution, environmental adapta- lana (Jalisco), A. deserti (the Sonoran Desert), and A. sisalana tion, and improvement of economically important traits. (Chiapas) [4, 6, 7]. Agaves have very large genomes, which has significantly restricted their genome assembly and lim- 2. Materials and Methods ited our understanding to their evolution pattern [16]. In other crops, accessible genomes and genome-wide associa- 2.1. Phylogenetic Analysis. Phylogenetic analysis was con- tion analysis have revealed many economically important ducted by MEGA 5.0 software with the minimum-evolution traits for crop improvement [17–19]. Recently, the devel- method [25]. The methods and parameters were according opment of next-generation sequencing has brought a new to the previous study [15]. The bootstrap method was direction for gene-related studies without the restriction employed for confidence in nodes with 1000 replicates. Partial of genome data [20, 21]. Furthermore, transcriptome chloroplast sequences for A. tequilana and A. deserti were International Journal of Genomics 3 A. tequilana A. sisalana 7000 6000 2,864 5000 13,452 38,721 4000 6,130 3000 6,939 3,154 2000 Number of unigene 1000 13,104 0 A. deserti 70 75 80 85 90 95 100 Te sequence identity (%) A. sisalana vs A. tequilana A. sisalana vs A. deserti A. tequilana vs A. deserti (a) (b) Figure 2: Sequence comparisons between domesticated A. tequilana and A. sisalana and wild A. deserti transcriptomes. (a) The sketch map showing 6130 unigene terms that were identified within the three transcriptomes. (b) The identity distribution of all orthologous unigene pairs. downloaded from NCBI (https://www.ncbi.nlm.nih.gov/) with 2.4. In Silico Gene Expression Analysis. The expression pat- the accession numbers GAHU01110124 and GAHT01022741, tern of positive selected unigenes was subjected to in silico respectively. Partial chloroplast sequences for A. sisalana gene expression analysis in agave leaves. The reads per kilo- were obtained from a previous transcriptome database base per million mapped read (RPKM) value of these uni- [24]. The whole chloroplast sequence of A. americana genes in A. deserti and A. tequilana were calculated by was obtained from NCBI under the accession number RSEM software according to the previous study [23, 30]. KX519714. The trnL+trnL-trnF sequences (about 900 bp) The RPKM data was further normalized with two reference for the 4 and other 14 agave species were also downloaded genes (tubulin and serine/threonine-protein phosphatase) according to the previous study [15]. in each agave species [31]. 2.5. Selection Pressure Detection and Protein Structure 2.2. Transcriptome Data and Gene Annotation. Three agave Modeling. To detect the selection pressure on positive selec- transcriptome databases were according to previous studies tion unigenes, Ka/Ks ratios were calculated in sliding window [23, 24]. Transcriptome assembly of the Illumina
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