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Frontiers of Agricultural Science and Engineering

ISSN 2095-7505

ISSN 2095-977X(Online)

CN 10-1204/S

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Front. Agr. Sci. Eng.    2021, Vol. 8 Issue (2) : 280-291    https://doi.org/10.15302/J-FASE-2020374
RESEARCH ARTICLE
FUNCTIONAL GAIN OF FRUIT NETTED-CRACKING IN AN INTROGRESSION LINE OF TOMATO WITH HIGHER EXPRESSION OF THE FNC GENE
Chunli ZHANG, Taotao WANG, Jing LI, Danqiu ZHANG, Qingmin XIE, Shoaib MUNIR, Jie YE, Hanxia LI, Yongen LU, Changxian YANG, Bo OUYANG, Yuyang ZHANG, Junhong ZHANG, Zhibiao YE()
Key Laboratory of Horticultural Plant Biology (Ministry of Education), Huazhong Agricultural University, Wuhan 430070, China
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Abstract

• A novel netted-cracking fruit phenotype was discovered in tomato introgression line IL4-4.

• A single dominant gene (FNC) determined the fruit netted-cracking phenotype.

• The high transcript level of FNC results in the functional gain of fruit netted-cracking and it was found to be a common mechanism in a diverse range of plant species.

Fruit cracking is a major disorder that affects the integrity of fruit and reduces the commercial value of tomato and other fleshy fruit. Here, we have found a novel fruit ‘netted-cracking’ (FNC) phenotype in tomato introgression line IL4-4 which is present in neither the donor parent (LA0716) nor the receptor parent (M82). An F2 population was generated by crossing IL4-4 with M82 to genetically characterize the FNC gene and this showed that a single dominant gene determined fruit netted-cracking. Further map-based cloning narrowed down the FNC locus to a 230 kb region on chromosome 4. Sequencing and annotation analysis show that FNC (Solyc04 g082540) was the most likely candidate gene. Functional characterization of FNC by overexpressing FNCAC and FNCIL4-4 resulted in the fruit netted-cracking phenotype, suggesting that the FNC transcript level results in the functional gain of fruit netted-cracking. These findings were further confirmed by FNC ortholog in netted-cracking pepper and melon, indicating a common regulatory mechanism in different plant species. Furthermore, cytoplasm and nucleus-localized FNC indicates increased expression of genes involved in suberin, lignin, lipid transport and cell wall metabolism. These findings provide novel genetic insights into fruit netted-cracking and offer a way to promote molecular improvement toward cracking resistant cultivars.
Keywords fine mapping      fruit netted-cracking      introgression line      transcript level     
Corresponding Author(s): Zhibiao YE   
Just Accepted Date: 31 December 2020   Online First Date: 11 February 2021    Issue Date: 13 July 2021
 Cite this article:   
Chunli ZHANG,Taotao WANG,Jing LI, et al. FUNCTIONAL GAIN OF FRUIT NETTED-CRACKING IN AN INTROGRESSION LINE OF TOMATO WITH HIGHER EXPRESSION OF THE FNC GENE[J]. Front. Agr. Sci. Eng. , 2021, 8(2): 280-291.
 URL:  
https://academic.hep.com.cn/fase/EN/10.15302/J-FASE-2020374
https://academic.hep.com.cn/fase/EN/Y2021/V8/I2/280
Fig.1  Tomato fruit netted-cracking phenotypes (red arrow) of IL4-4. (a) The fruit cracking of different ripening stages at immature (IM), breaker (BR) and red ripe (RR) in IL4-4 compared with M82. (b) The netted-cracking structures of pericarps in IL4-4 and M82 fruit at 14, 21, 28 and 35 days post anthesis (DPA). (c) The ultrastructures of cracking tissues in IL4-4 and M82 fruit at 21, 28 and 35 DPA observed under scanning electron microscope. (d) Constructing F2 population from selfing F1 crossed with M82 and IL4-4.
Fig.2  Fine mapping of the fruit netted-cracking gene (FNC) of tomato. (a) The locus of netted-cracking phenotype is located in the specific region of IL4-4. (b,c) Mapping the FNC gene to the telomeric portion of chromosome 4 between the markers of T0974 and TG163 (b) and further mapped in the specific region of IL4-4 between M27 and M28 (c). (d,e) The candidate interval determined between markers M47 and M19 (d), and the physical location of the genome at 63.75–63.98 Mb, and the localized candidate FNC gene is Solyc04 g082540 (e).
Fig.3  FNC gene expression and evolution analyses. (a) The analysis of conserved domain of the FNC gene. (b) The expression of the FNC gene in IL4-4 and M82 at different days post anthesis (DPA). (c) Phylogenetic analysis of the FNC gene. Diverse colors represent different species, FNC and potato PGSC0003DMT400025661 are classified into one group (yellow).
Fig.4  Overexpressing the FNC gene in tomato leads to the fruit netted-cracking phenotype. (a) FNCIL4-4 protein has three more b-fold structures than FNCAC. Red (FNCIL4-4), Blue (FNCAC). The yellow arrows indicate three of the b-fold structures. (b) The transcript levels of overexpressing FNCIL4-4 and FNCAC, Ailsa Craig (AC) as a control cultivar, (OE) overexpression. (c) Overexpression of FNCIL4-4 and FNCAC in a non-cracking AC resulted in a netted-cracking on the fruit surface.
Fig.5  Subcellular localization of the FNC protein observed by fluorescence confocal microscopy. The transient expressions of 35S::YFP (a) as control and 35S::FNC-YFP (b) in tobacco leaves.
Fig.6  Increased expression of the FNC gene can affect the metabolic pathway of pericarp of tomato. (a–e) The relative expression genes involved in various metabolism pathways such as in suberin metabolism pathway (a), in lignin metabolism pathway (b), in lipid metabolism pathway (c), in lipid transport pathway (d) and in cell wall metabolism pathway (e) in FNC-OE plants and AC. SlGPAT5 (Solyc04 g011600), SlASFT (Solyc03 g097500), PAL (phenylalanine ammonia lyase, Solyc10 g086180), 4CL (4-coumaric acid, CoA ligase, Solyc06 g068650), Laccase-13 (laccase, Solyc06 g082240), GDSL lipase, KCS (b-ketoacyl CoA synthase), FAR (fatty acyl reductase), NLTP9 (Solyc09 g082270), LTP5 (Solyc09 g065430), EXPA11 (Solyc04 g081870) and PG (Solyc08 g060970). (f) The ultrastructure of fruit epidermis in the overexpressing plants and the control cultivar during the breaker stage.
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