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. 2021 Sep 30;10(10):2062.
doi: 10.3390/plants10102062.

Genetic Mapping of the HLA1 Locus Causing Hybrid Lethality in Nicotiana Interspecific Hybrids

Takahiro Tezuka  1   2 , Naoto Kitamura  1 , Sae Imagawa  3 , Akira Hasegawa  3 , Kumpei Shiragaki  1 , Hai He  1 , Masanori Yanase  1 , Yoshiyuki Ogata  1 , Toshinobu Morikawa  1 , Shuji Yokoi  1   2   4
Affiliations

Genetic Mapping of the HLA1 Locus Causing Hybrid Lethality in Nicotiana Interspecific Hybrids

Takahiro Tezuka et al. Plants (Basel). .

Abstract

Hybrid lethality, a postzygotic mechanism of reproductive isolation, is a phenomenon that causes the death of F1 hybrid seedlings. Hybrid lethality is generally caused by the epistatic interaction of two or more loci. In the genus Nicotiana, N. debneyi has the dominant allele Hla1-1 at the HLA1 locus that causes hybrid lethality in F1 hybrid seedlings by interaction with N. tabacum allele(s). Here, we mapped the HLA1 locus using the F2 population segregating for the Hla1-1 allele derived from the interspecific cross between N. debneyi and N. fragrans. To map HLA1, several DNA markers including random amplified polymorphic DNA, amplified fragment length polymorphism, and simple sequence repeat markers, were used. Additionally, DNA markers were developed based on disease resistance gene homologs identified from the genome sequence of N. benthamiana. Linkage analysis revealed that HLA1 was located between two cleaved amplified polymorphic sequence markers Nb14-CAPS and NbRGH1-CAPS at a distance of 10.8 and 10.9 cM, respectively. The distance between these markers was equivalent to a 682 kb interval in the genome sequence of N. benthamiana.

Keywords: Nicotiana debneyi; Nicotiana fragrans; hybrid lethality; interspecific population; linkage analysis; reproductive isolation.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
RAPD, AFLP, and SSR markers linked to the HLA1 locus. (a,b) RAPD patterns amplified with primers OPA-16 (a) and OPA-19 (b). (c,d) AFLP patterns amplified with primers E40 and M58 (c), and primers E37 and M52 (d). (e) SSR pattern amplified with primers for PT30138 marker. Lanes: M, size marker (GeneRuler DNA ladder mix; Thermo Fisher Scientific, Tokyo, Japan); 1, N. debneyi; 2, F1 hybrid between N. debneyi and N. fragrans; 3, N. fragrans. Black and white arrowheads indicate dominant markers specific to N. debneyi and N. fragrans, respectively. Black and white arrows indicate codominant marker PT30138a bands specific to N. debneyi and N. fragrans, respectively.
Figure 2
Figure 2
DNA markers developed based on v0.4.4 genome of N. benthamiana. (a–c) CAPS markers NbRGH1-CAPS (a) and NbRGH3-CAPS (c), and dCAPS marker NbRGH2-dCAPS (b). Undigested PCR products (lanes 1–3) and PCR products treated with appropriate restriction enzymes (lanes 4–6) were detected by agarose gel electrophoresis. (d) STS marker NbRGH4 detected by polyacrylamide gel electrophoresis. Lanes: M, size marker (GeneRuler DNA ladder mix); 1, 4, N. debneyi; 2, 5, F1 hybrid between N. debneyi and N. fragrans; 3, 6, N. fragrans.
Figure 3
Figure 3
Linkage map (left) and physical map (right) indicating the position of the HLA1 locus. The physical map was based on the v1.0.1 genome of N. benthamiana.
Figure 4
Figure 4
DNA markers developed based on the scaffold Niben101Scf06736 in the v1.0.1 genome of N. benthamiana. (a) STS marker Nb7 (arrowhead). (b,c) CAPS marker Nb14-CAPS. Undigested PCR products (b) and PCR products treated with HhaI (c) are shown. Lane M, size marker (GeneRuler DNA ladder mix).
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