Assessment of genetic diversity and chemical composition among seven black locust populations from Northern China

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Highlights

  • FX population of black locust has the potential to be a source of lignocellulosic biomass for bioenergy.

  • 14 microsatellite markers are successfully used to evaluate the genetic diversity of black locust populations.

  • High genetic diversity and low genetic differentiation exist among seven black locust populations.

  • Chemical composition has no relationship with genetic variation and geographic distribution in black locust populations.

Abstract

Black locust (Robinia pseudoacacia L.) is one of the important biomass sources used to produce bioenergy and bioethanol. In this study, we examined variations in chemical composition as well as genetic diversity and differentiation among 165 black locust plants from seven populations in five provinces (Beijing, Hebei, Gansu, Shanxi, Shandong) of Northern China using microsatellite markers(SSR). The contents of cellulose, hemicellulose, lignin varied widely among seven populations. Of the microsatellite markers analysed, 14 showed polymorphisms, and 45 alleles were identified. The polymorphism information content (PIC) ranged from 0.2885 (Rops4) to 0.6837 (Rp200), and most of the microsatellite loci had PIC values > 0.5. Expected heterozygosity (He), observed heterozygosity (Ho), and Shannon's information index (I) detected relatively high genetic variation among populations. The percentage of polymorphic loci in three populations was 100%, and the average among all populations were 95.92%. Analysis of molecular variance (AMOVA) indicated that the degree of genetic differentiation among the seven populations was low (GST = 0.058; Nm = 4.05), and chemical compositions had no relationship with genetic or geographic distance. This study demonstrates that microsatellite markers efficiently assess of genetic diversity and genetic differentiation in black locust populations, and all seven populations exhibit high genetic diversity. The population from Feixian has the potential to be lingo-cellulosic biomass for bioenergy and bioethanol.

Introduction

The black locust (Robinia pseudoacacia. L), a member of the Fabaceae family, has several prominent characteristics including nitrogen fixation, stress tolerance, fast growth, high-density wood, and carbon sequestration (Straker et al., 2015). The black locust is an introduction species, which originated from a small area of the Appalachian region in the south-eastern United States. The black locust was one of the earliest species introduced into Europe from North America in the early 17th century, and it was later imported from Europe to Shandong province of China in the 1880s and 1890s. Nowadays, the black locust is one of the most widely cultivated and important woody species in China (Vítková et al., 2017). It can be widely to achieve the purpose of industry (mining, construction, and furniture), apiculture and honey production, reforestation, biodiversity and soil and water conservation (Redei et al., 2008). As an important short-rotation woody crop, black locust can be coppiced (Rédei et al., 2010), so it produces large amounts of woody biomass. Thus, humans use black locust as a woody energy biomass in combined heat and power (CHP) systems, and bioethanol or biofuel can be extracted from cellulose of black locust (Balat, 2010; Monedero et al., 2017). Short-rotation forestry cultivation and life-cycle assessment have been employed to obtain large amounts of cellulose as feedstock for ethanol production from black locust trees (González et al., 2011, 2012). In summary, this species show strong potential as a feedstock for renewable bioenergy (Grünewald et al., 2009).

The biofuel and bioethanol from biomass increased during the recent decades. Bioenergy, as a renewable energy is being considered as one way to solve the problem of energy security and environmental safety. However, the shortage of non-food feedstock is a most important problem for the production of bioenergy. The corn, sugarcane, rice and wheat, straws are the most main cellulosic feedstock in the United States, South America, Asia, and Europe, respectively (Limayem and Ricke, 2012). Lignocellulose can be obtained from non-food sources, as an abundant feedstock, it can be used to produce bioethanol. The lignocellulose composes of three main components: cellulose which consists of long chains of β-glucose monomers (30–50%); hemicellulose which compose of mainly pentose (such as xylose and arabinose) and hexoses (such as mannose, glucose, and galactose)(15–35%) (Gírio et al., 2010; Rastogi and Shrivastava, 2017); It can be seen that cellulose and hemicellulose are the main feedstock in the transformation of lingo-cellulosic biomass into bioethanol (Aditiya et al., 2016; Haghighi et al., 2013; Limayem and Ricke, 2012; Zabed et al., 2017). Face to the shortage of feedstock for production of bioenergy, there are many barren hills that can be used to planting the black locust in China and provided a large amount of feedstock for bioenergy.

Genetic diversity and differentiation can be evaluated at different levels, including morphological, cytological, physiological, biochemical, or DNA markers. Molecular markers based on DNA have developed and matured for genetic and ecology. These techniques include random amplification of polymorphic DNA (RAPD) (Bindiya and Kanwar, 2003; Filippis and Magel, 2002; Shu et al., 2003), amplified fragment length polymorphism (AFLP) (Li et al., 2008; Pecina et al., 2013), and inter-simple sequence repeat (ISSR) analyses (Sharma et al., 2016; Sun et al., 2018). Simple sequence repeat (SSR) markers become powerful molecular tools for assessing genetic variation due to the characteristics of high information content, polymorphism, co-dominant inheritance and low-cost. In the past, genetic diversity and population variation in black locust were only studied based on molecular marker approaches such as analyses of allozymes, RAPD, AFLP markers, and sequence-related amplified polymorphism (SRAP) (Gu et al., 2010; Xuemei et al., 2009; Yang et al., 2004; Yuan et al., 2012). Although, microsatellites markers have been developed for the black locust so that they can provide a marker system for genetic diversity and solve issue of breeding programs and germplasm resource utilisation, that only be used to assess genetic diversity and differentiation of black locust seeds which endured 15days in spaceflight Yuan et al. (2012). Few studies use both molecular markers and chemical characteristics to examine population variation in the black locust.

In this paper, we used the microsatellites markers and advanced chromatography techniques to examine the genetic variability and chemical components of black locust populations from different regions in northern China, respectively. The main objectives of this study are follows as: (1) estimate chemical variation in different populations of black locust in Northern China; (2) determine the genetic diversity and genetic differentiation among seven populations using microsatellites markers; (3) analyse possible associations of chemical composition with genetic distance and geographic location to identify superior genotypes. Finally, we find potential source of cellulosic biomass for bioenergy and bioethanol, and provide scientific guidance for improving germplasm resources of black locust.

Section snippets

Collection of plant materials

We collected 165 individuals from seven populations in five different provinces (Hebei, Shanxi, Shandong, Gansu, and Beijing) of northern China (Fig. 1). The locations and environmental parameters of the seven black locust populations are listed in Table 1. The black locust populations included four locations where the quality of black locust seedlings was supervised by the State Forestry Bureau of China. Each collected plant was more than 50 m away from others. The black locust scions were

Chemical compositions

The chemical compositions of the seven black locust populations are listed in Table 3. All of the chemical composition showed significant differences (P < 0.05) except the content of arabinose. The cellulose content ranged from 31.53 ± 2.96% (HY) to 37.63 ± 3.39% (FX), and the FX population was significantly higher than other populations; the hemicellulose content ranged from 15.06 ± 1.23%(YQ) to 19.02 ± 2.52%(FX) comprised of four pentose with content ranges of 8.67 ± 0.75–12.23 ± 0.78%

Discussion

Cellulose is the mainly important feedstock in the bioenergy and bioethanol. Thus, the first aim is to determine the diversity of chemical components among black locust populations from different regions in this study. Cellulose contents in the FX populations were significantly higher than others. Furthermore, based on Duncan's multiple range test, the FX population had the highest hemicellulose and xylopyranose contents. Overall, black locust trees from the FX population have the best

Conclusion

From the perspective of the objectives in this study, there were significant variations in the chemical composition and molecular markers among the seven black locust populations in north China. The black locust population from Feixian had the highest content of cellulose, which made it have the potential to be biomass for lingo-cellulosic bioenergy and bioethanol. 14 microsatellite markers were proven to be useful tools for exploring genetic diversity and differentiation. According to genetic

Author contributions

Xinchao Yang designed the experiment, analyzed the data and wrote the manuscript, Yun Li, Jie Duan collected and cultivated the plant materials. Xinchao Yang, Kaiquan Zhang, Hansen Jia, and Jing Wang worked the experimental study. Lvyi Ma carried out data correction and manuscript revision.

CRediT authorship contribution statement

Xinchao Yang: Conceptualization, Formal analysis, Investigation, Writing - original draft, Writing - review & editing. Kaiquan Zhang: Data curation, Investigation. Jing Wang: Writing - review & editing, Investigation. Hansen Jia: Investigation. Lvyi Ma: Conceptualization, Writing - review & editing, Funding acquisition. Yun Li: Resources, Funding acquisition. Jie Duan: Project administration.

Declaration of competing interest

The authors report no conflicts of interest.

Acknowledgments

This work was financial supported by the National Key R&D Program of China (2017YFD0600503), and the International S&T Cooperation Program of China (2014DFA31140). We thank Director Shaoming Wang (Lyucun forest farm, Luoning, Henan province) for assistance in grafted and planted the black locust experimental material.

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