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Research Article

Genetic Characterization of BADH2 in Philippine Aromatic Rice Cultivars

Plant Breeding and Biotechnology 2021;9(3):227-238.
Published online: September 1, 2021

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*Corresponding author Marjohn C. Niño, marjohn.nino@ctu.edu.ph, Tel: +63-32-513-0641, Fax: +63-32-412-0970
• Received: July 14, 2021   • Revised: August 4, 2021   • Accepted: August 4, 2021

Copyright © 2021 by the Korean Society of Breeding Science

This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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  • Variability in 2-acetyl-1-pyrroline production and associated mutations in BADH2 gene in aromatic rice cultivars of Bangladesh
    Mahmuda Umme Rayhan, Habibul Bari Shozib, Fardous Mohammad Safiul Azam, Tofazzal Islam
    Gene Reports.2023; 33: 101847.     CrossRef

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Genetic Characterization of BADH2 in Philippine Aromatic Rice Cultivars
Plant Breed. Biotech.. 2021;9(3):227-238.   Published online September 1, 2021
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Genetic Characterization of BADH2 in Philippine Aromatic Rice Cultivars
Plant Breed. Biotech.. 2021;9(3):227-238.   Published online September 1, 2021
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Genetic Characterization of BADH2 in Philippine Aromatic Rice Cultivars
Image Image Image Image
Fig. 1 PCR banding pattern of 28 cultivars using markers designed by (A) (Bradbury et al.. 2005) and (B) (Shi et al. 2008). In (A), all samples have a common product with a length of approximately 580 bp. Cultivars with an 8-bp deletion and 3 SNPs in exon 7 had a second product of 257 bp in size, whereas cultivars without the deletion and SNP in exon 7 had a second product of 355 bp in size. However, in (B), the 7-bp deletion in exon 2 (badh2.2) was not detected as all cultivars showed bands that are similar to the negative checks.
Fig. 2 Multiple sequence alignment of BADH2 gene exons 1, 2, 7, 8, 9, 10, 11, 12, 14, and 15 in selected cultivars revealing some SNPs and deletions. The sequences were aligned with BAC clone AP005537.3 and LOC_Os08g 32870. (A) 109-bp long exon 1 of BADH2 gene; (B) 143-bp long exon 2; (C) 67-bp long exon 7 revealing badh2.1 allele showing the mutations at position 4230-4237 flanked by 3 SNPs at position 4226, 4228, and 4238 in Azucena, Burdagol, KDML 105, and Sampaguita; (D) 67-bp long exon 8; (E) 77-bp long exon 9; (F) 114-bp long exon 10; (G) 70-bp long exon 11; (H) 105-bp long exon 12; (I) 107-bp long exon 14; and 91-bp long exon 15.
Fig. 3 An unrooted neighbor-joining tree showing the relationships of nine cultivars sequenced at the BADH2 region and Nipponbare. Taxa with shaded circles are aromatic (based on prior information), taxon with open circle (Nipponbare) is non-aromatic.
Fig. 4 The relative transcript abundance expressed as relative expression ratio (rER) using RT-qPCR (gray bar) and the 2-acetyl-1-pyrroline (2AP) levels in 22 cultivars cultivated during wet season (blue bars) and dry season (orange bars). Error bars represent the standard deviation determined from three replicates.
Genetic Characterization of BADH2 in Philippine Aromatic Rice Cultivars

List of the rice cultivars used in this study and their corresponding attributes.

Cultivar Origin Attributes IRGC accession number
Nipponbare (Nb) Japan Nonaromatic, negative control IRGC 117274
IR64 IRRI Nonaromatic, negative control IRGC 66970
Khao Dawk Mali 105 (Kd) Thailand Aromatic, positive control IRGC 27748
Basmati 370 (Bm) India and Pakistan Aromatic IRGC 4895
Dom-sofid (Ds) Iran Aromatic IRGC 12880
Gaen Magawk (Gm) Vietnam Aromatic IRGC 7269
Khao Intok (Ki) Thailand Aromatic IRGC 12951
IR24 IRRI Nonaromatic
NSIC Rc134 (PJ21) PhilRice-JICA Nonaromatic
NSIC Rc146 (PJ7) PhilRice-JICA Nonaromatic
Azucena (Az) Philippines Aromatic, traditional IRGC 328
Binaka (Bk) Philippines Aromatic, traditional
Burdagol (Bd) Philippines Aromatic, traditional
Dinalores (Dl) Philippines Aromatic, traditional
Dinorado (Dn) Philippines Aromatic, traditional
Finongod (Fn) Philippines Aromatic, traditional
Laila (Ll) Philippines Aromatic, traditional IRGC 34359
Macaraniag (Mc) Philippines Aromatic, traditional IRGC 44570
Mimis (Mm) Philippines Aromatic, traditional
Minantika (Mn) Philippines Aromatic, traditional
Minerva (Mv) Philippines Aromatic, traditional
Perurutong Magdalena (PM) Philippines Aromatic, traditional
Saigorot (Sg) Philippines Aromatic, traditional IRGC 44727
Salanay (Sy) Philippines Aromatic, traditional
Salumpikit (Sl) Philippines Aromatic, traditional
Sampaguita (Sp) Philippines Aromatic, traditional
Silimut (St) Philippines Aromatic, traditional IRGC 43621
Wagwag Los Baños (Ww) Philippines Aromatic, traditional IRGC 44803

The estimates of evolutionary divergence between BADH2 gene sequences of selected cultivars. Distances (d) (number of substitutions per site) based on Maximum Composite Likelihood model are shown below the diagonal, while standard error estimate(s) are shown above the diagonal. The average divergence value of a cultivar to the rest of the group is also indicated.

Nb Az Bd Kd Mm Mv Sm Dn Sg Sy Ave (d)
Nb 0.0007 0.0008 0.0007 0.0008 0.0004 0.0008 0.0007 0.0013 0.0016 0.0038
Az 0.0018 0.0002 0.0002 0.0007 0.0005 0.0000 0.0009 0.0016 0.0017 0.0034
Bd 0.0021 0.0002 0.0003 0.0006 0.0006 0.0003 0.0009 0.0013 0.0016 0.0034
Kd 0.0023 0.0002 0.0005 0.0005 0.0007 0.0004 0.0009 0.0012 0.0015 0.0033
Mm 0.0027 0.0015 0.0023 0.0016 0.0009 0.0009 0.0008 0.0007 0.0009 0.0026
Mv 0.0006 0.0011 0.0010 0.0017 0.0026 0.0005 0.0008 0.0016 0.0019 0.0036
Sm 0.0022 0.0000 0.0007 0.0009 0.0024 0.0013 0.0008 0.0014 0.0014 0.0035
Dn 0.0026 0.0032 0.0034 0.0035 0.0029 0.0021 0.0034 0.0005 0.0006 0.0026
Sg 0.0091 0.0106 0.0092 0.0090 0.0031 0.0101 0.0095 0.0010 0.0009 0.0072
Sy 0.0106 0.0118 0.0107 0.0104 0.0040 0.0116 0.0108 0.0017 0.0032 0.0083

Analysis of variance for rER based on the empirical distribution function.

Source of variation Degrees of freedom Sum of squares Mean square F value Pr (>F)
Replication 2 0.010 0.0051 0.653 0.525
Cultivar 22 17.960 0.8164 105.441 <2E-16
Error 44 0.341 0.0077
Total 68 18.311
Table 1 List of the rice cultivars used in this study and their corresponding attributes.
Table 2 The estimates of evolutionary divergence between BADH2 gene sequences of selected cultivars. Distances (d) (number of substitutions per site) based on Maximum Composite Likelihood model are shown below the diagonal, while standard error estimate(s) are shown above the diagonal. The average divergence value of a cultivar to the rest of the group is also indicated.
Table 3 Analysis of variance for rER based on the empirical distribution function.