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Article

Morphological and Molecular Characterizations of Musa (ABB) ‘Mali-Ong’ in Thailand

by
Duangporn Premjet
1,2,
Thanita Boonsrangsom
1,2,
Kawee Sujipuli
1,2,
Kumrop Rattanasut
1,2,
Anupan Kongbungkerd
1,3 and
Siripong Premjet
1,3,*
1
Center of Excellence in Research for Agricultural Biotechnology, Faculty of Agriculture, Natural Resources and Environment, Naresuan University, Muang, Phitsanulok 65000, Thailand
2
Department of Agricultural Science, Faculty of Agriculture, Natural Resources and Environment, Naresuan University, Muang, Phitsanulok 65000, Thailand
3
Department of Biology, Faculty of Science, Naresuan University, Muang, Phitsanulok 65000, Thailand
*
Author to whom correspondence should be addressed.
Biology 2022, 11(10), 1429; https://doi.org/10.3390/biology11101429
Submission received: 27 August 2022 / Revised: 24 September 2022 / Accepted: 27 September 2022 / Published: 29 September 2022
(This article belongs to the Section Conservation Biology and Biodiversity)

Abstract

:

Simple Summary

The banana cultivar Musa (ABB) ‘Mali-Ong’ is widely used in the processed food industry in Thailand, where it determines the quality of the products. However, the sub-cultivars of ‘Nam Wa Mali-Ong’ are almost indistinguishable, with few morphological differences and minimal genetic variation. This study used 77 morphological characteristics and two types of molecular markers to distinguish Nam Wa Mali-Ong from other cultivars. The study also assessed the genetic variation of nine ‘Nam Wa Mali-Ong’ clones and compared them with 10 other samples of bananas with different genomes or chromosome sets. The molecular markers grouped the ‘Nam Wa Mali-Ong’ samples. Four clones (A, B, D, and I) were superior and had higher bunch weights. This study will be useful for germplasm evaluation and future ‘Nam Wa Mali-Ong’ improvements.

Abstract

Musa (ABB) ‘Mali-Ong’ is an economically important banana cultivar in Thailand. We morphologically and molecularly characterized ‘Nam Wa Mali-Ong’. Leaf blade width was the only statistically different morphological character among the clones. To determine genetic variation, nine ‘Nam Wa Mali-Ong’ clones were compared with 10 samples of Musa ABB, AA, and BB cultivars by fingerprinting using seven pairs of sequence-related amplified polymorphism (SRAP) and eight inter simple sequence repeat (ISSR) markers. The SRAP and ISSR primers generated 65 and 76 amplicons, respectively, of which 57 (87.7%) and 62 (81.6%) amplicons, respectively, were polymorphic; the polymorphic information content was 0.28–0.49. The SRAP data revealed two distinct groups: Group I, comprising two subgroups (one including all ABB samples and the other containing the BB genome accessions), and Group II, comprising the AA genome accessions. The ISSR data revealed two groups: Group I, which incorporated the AA (Hom Champa) genome, and Group II, consisting of two subgroups: Subgroup A, comprising only the AA (Hom Chan) accessions, and subgroup B, comprising all the ABB accessions and wild banana M. balbisiana (BB genome). The ‘Nam Wa Mali-Ong’ samples clustered together, regardless of the markers used. SRAP and ISSR markers will be useful for germplasm evaluation and future Musa (ABB) improvements.

1. Introduction

Banana (Musa sp.) is an economically important fruit grown in several tropical countries with a high demand in global markets. In 2021, approximately 100 million tons of bananas were supplied from across over 130 tropical nations [1]. Banana has a high nutritional value and numerous health benefits. It is rich in dietary fiber, potassium, antioxidants, vitamins B6 and C, and phytonutrients. Bananas are widely consumed fresh and processed to produce sun-dried bananas, banana flour, banana starch, and various snacks [2]. Edible banana cultivars are almost exclusively the result of interspecific crosses between two wild species. A cross between Musa acuminata (AA genome) and Musa balbisiana (BB genome) produces the AAB, ABB, AABB, and ABBB genome groups. The triploids (AAA and AAB cultivars) are cultivated globally and play a significant role in the global economy [3]. The ABB cultivar originated in South-East Asia and India and is divided into nine subgroups: Bluggo, Monthan, Ney Mannan, Pisang Awak, Peyan, Pelipita, Saba, Kalapua, and Kloe Teparod [4]. In Thailand, banana (especially the ABB cultivar) is the most important fruit crop because of its high biotic and abiotic stress resistance. Moreover, bananas contain polyphenols that can act as health-promoting substances and exhibit an anti-diabetic effect [5]. Many banana cultivars with ABB genomes, such as ‘Kluai Tip Yai’, ‘Kluai Hak Muk’, and ‘Kluai Nam Wa,’ are grown in Thailand. The ‘Kluai Nam Wa’ cultivar is extremely valuable to the banana processing business [6]. It is widely planted in northern Thailand, especially in the provinces of Phitsanulok and Nan. Kluai Nam Wa is sweet and can be eaten fresh without heat treatment [7]. The fruits are nutritious and processed as high-quality sundried bananas, with a yellowish-brown color, sweet flavor, and soft texture. Many sub-cultivars, such as Kluai Nam Wa Deang, ‘Kluai Nam Wa Khwan’, and ‘Kluai Nam Wa Mali-Ong’, belong to the Musa (ABB genomes) complex. These sub-cultivars are almost indistinguishable, with few morphological differences and minimal genetic variation. Therefore, identifying the ‘Kluai Nam Wa’ sub-cultivars remains difficult. To date, scarce scientific information is available on the conservation and utilization of ‘Kluai Nam Wa Mali-Ong’ and its morphology and phylogenetic relationship. The definition of genetic variation or cultivar is the foundational step in germplasm management, not only for identifying features but also for facilitating their suitable application [8]. Characterization is suggested using morphotaxonomy, cytology, and molecular genotyping, including of nutritional traits [9]. Molecular markers, including random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR), simple sequence repeat (ISSR), and sequence-related amplified polymorphism (SRAP) markers, have been employed to examine genetic diversity among banana accessions and species [2,10,11,12]. SRAP markers have been used to determine the relationship of a cross between Musa (ABB) and two wild cultivars (AA and BB) [13]. However, no precise information exists on the morphological traits of ‘Kluai Nam Wa Mali-Ong’. This cultivar is the most important banana for the food industry in Thailand and determines the quality of the products. The collection of clones distributed throughout Phitsanulok in northern Thailand is essential for establishing ex situ conservation of ‘Kluai Nam Wa Mali-Ong’ to preserve an elite cultivar that will strengthen and sustainably drive the bio, circular, and green economy. The objectives of this study were: (1) to collect nine ‘Kluai Nam Wa Mali-Ong’ clones from a farm in Phitsanulok to describe their morphological characteristics; and (2) to evaluate the genetic variation of ‘Kluai Nam Wa Mali-Ong’ collected from agricultural fields in nine districts of Phitsanulok Province in Thailand using SRAP and ISSR markers.

2. Materials and Methods

2.1. Collection of Plant Materials

Ten sword suckers of Musa (ABB) ‘Mali-Ong’ were collected in February 2021 from a farm in each of the 12 plantation areas in nine districts of Phitsanulok Province: Muang, Bang Rakam, Bang Kratum, Nakhon Thai, Chat Trakarn, Wang Thong, Neon-Maprang, Wat Bot, and Prom Phiram (Figure 1). Musa reference genomes (AA, BB, and ABB) were obtained from the banana germplasm at the Nakhon Ratchasima and Sukhothai Horticultural Research Center. Banana samples were coded as shown in Table 1.

2.2. Observation of Agronomic Traits

‘Kluai Nam Wa Mali-Ong’ accessions were cultivated with a 4 × 4 m spacing in a field at Plant Propagation Center No. 6, Phitsanulok, Thailand. The banana plants were provided with 17–20 L of water per day using a drip water system. Compost (1 kg plant−1) was applied once a month for 7 months, and chemical fertilizer (15-15-15) was applied 7 months after transplantation at a rate of 125 g plant−1. The bunch weight (kg) and the number of hands/bunch were recorded.

2.3. Voucher Deposition of Musa (ABB) ‘Mali-Ong’

The herbarium in the Department of Biology, Naresuan University, confirmed the botanical identity of the plants. Samples of Musa (ABB) ‘Mali-Ong’ were deposited at the Naresuan University and Laboratory Herbarium (herbarium deposit vouchers 05780 and 05781) for future reference.

2.4. Determination of Morphological Characteristics

We determined the morphological parameters of Musa (ABB) ‘Mali-Ong’ cultivated under standardized conditions from 8–16 months (plant to fruit). A morphological record was made according to the 1996 Book of Descriptors for Banana (Musa spp.) [14]. The following 77 characteristics were recorded: 7 features of the artificial trunk (pseudostem), 17 of the leaf, 9 of the inflorescence/plant (male bud), 8 of the bracts, 16 of the male flowers, and 20 of the fruit. The Royal Horticultural Society Color Charts were used to specify colors. Three samples of each clone were evaluated for their qualitative and quantitative characteristics. The ‘Nam Wa Mali-Ong’clones were named as follows: clone A, Muang; clone B, Wang Thong; clone C, Bang Rakam; clone D, Wat Bot; clone E, Bang Kratum; clone F, Prom Phiram; clone G, Neon-Maprang; clone H, Nakhon Thai; and clone I, Chat Trakarn.

2.5. Genetic Variation Assessment Using Molecular Markers

2.5.1. Primers

Eight ISSR primers and seven SRAP marker pairs (Table 2 and Table 3) were purchased from Gibthai Co., Ltd. (Bangkok, Thailand).

2.5.2. Genomic DNA Extraction

Six months after planting, the cigar leaves of the sample plants were used for DNA extraction. Two samples were obtained from each collection site for DNA extraction. Genomic DNA was extracted using the PureDireX Genomic DNA Isolation Kit (Plant) (Bio-Helix Co., Ltd., New Taipei City, Taiwan) according to the manufacturer’s instructions. Briefly, 100 mg of banana leaves were pulverized in liquid nitrogen to a powdered form, and 500 µL of buffer PL and 2.5 µL of RNase A (10 mg mL−1) were added to the powder in 1.5 mL tubes and stirred gently. The tubes were incubated for 60 min at 75 °C and centrifuged at 14,462× g for 5 min. The clear top layer was transferred to a new 1.5 mL tube, and an equal amount of cold isopropanol was added. Subsequently, 400 µL of buffer W1 and the mixture were added to a column PC and centrifuged at 14,462× g for 30 s. The column PC was removed from its container and returned to the original collection tube after centrifugation. The centrifuged flowthrough was discarded. To retrieve DNA, 600 µL of W2 buffer (ethanol:buffer = 1:4) was poured into column PC and centrifuged for 2 min at 14,462× g. The DNA precipitate was preserved at −20 °C until further use.

2.5.3. DNA Quality and Quantity Determination

The quantity and purity of the DNA were measured using 0.8% agarose gel in a 1× TAE buffer and RedSafeTM nucleic acid staining solution. The intensity of the ultraviolet fluorescent bands was proportional to the amount of DNA. The intensity was compared with a known standard DNA ladder of 100 bp (ONEMARK 100 DNA bp ladder, Bio-Helix Co. Ltd., New Taipei City, Taiwan) [13].

2.5.4. ISSR-PCR

Twenty ISSR primers were tested, but only eight ISSR primers provided clear bands. These eight ISSR primers were used for polymerase chain reaction (PCR) amplification [11,15,16,17,18,19]. The 20 µL reaction mixture consisted of 10× PCR buffer, MgCl2 (0.2 mM), Taq polymerase (1.25 U), and 50 ng genomic DNA. A thermal cycler was used to perform DNA amplification (T100TM, Bio-Rad, Hercules, CA, USA). An initial denaturation at 94 °C for 5 min was followed by 35 cycles of denaturation for 30 s at 94 °C, annealing for 1 min at 50–53 °C, extension for 1 min at 72 °C, and a final extension for 2 min at 72 °C [13].

2.5.5. SRAP-PCR

Sixty-four pairs (eight forward primers: Me1, Me2, Me3, Me4, Me5, Me6, Me7, and Me9; and eight reverse primers: Em1, Em2, Em3, Em4, Em5, Em6, Em7, and Em8) were screened to generate clear DNA bands. A total of 20 µL of 2× OnePCRTM Plus (Bio-Helix Co., Ltd., New Taipei City, Taiwan), including Taq DNA polymerase, dNTP mix and gel loading dye, 5 M SRAP primers, and 50 ng of genomic DNA, was used for the PCR experiments. Thermal cycler amplifications were carried out using a denaturation step at 94 °C for 3 min, followed by 5 cycles of 3 steps: 1 min for denaturation at 94 °C, 1 min for annealing at 35 °C, and 2 min for an extension at 72 °C, followed by 35 cycles with an annealing temperature of 50 °C and a final prolongation step for 5 min at 72 °C [13].
The ISSR and SRAP-PCR amplicons were separated on 1.5% (w/v) agarose gel plates, stained with RedSafeTM, intron biotechnology, Inc., Gyeonggi, Korea and visualized using a gel documentation system (Thermo Fisher Scientific, Taoyuan City, Taiwan). The size of each fragment was estimated using the ONEMARK 100 DNA Ladder (Bio-Helix Co., Ltd., New Taipei City, Taiwan). Gel documentation systems were used to observe the ISSR and SRAP-PCR amplicons on an agarose gel stained with RedSafeTM, intron biotechnology, Inc., Gyeonggi, Korea. This DNA Ladder (ONEMARK 100) was used to assess the size of each of the fragments (Bio-Helix Co., Ltd., New Taipei City, Taiwan).

2.6. Analysis of Morphological Data

The mean and standard deviation were presented for the quantitative morphological parameters. Analysis of variance (ANOVA) was performed to examine whether the results from the nine clones were significant using a randomized plot design and one-way ANOVA. The DMRT test (p < 0.05) investigated significant differences between the mean values.

2.7. Molecular Data Analysis

To build a binary matrix, the SRAP and ISSR amplicons were scored as either 1 (present) or 0 (absent). Polymorphism (%) and the average number of bands per primer were computed from the total number of amplified bands. The polymorphic information content (PIC) was calculated using the following formula: PIC =   1 P i 2 , where Pi is the ith allele frequency for an individual population [20]. Free Tree & Tree View, a freeware tool, was used to build phylogenetic trees using the unweighted pair-group method with arithmetic average (UPGMA) analysis based on Dice similarity coefficients, and a bootstrap analysis based on 1000 bootstrap repeats was performed [21].

3. Results

3.1. Morphological Characteristics of Musa (ABB) ‘Mali-Ong’

The 77 morphological features are listed in Table 4 and shown in Figure 2. An intermediate leaf habit characterizes the overall appearance of a plant. ‘Nam Wa Mali-Ong’ has a tall pseudostem with an average height (326.8225 cm) > 3 m. The pseudostem is slender (60.767 cm), yellow-green, and waxy. There are eight suckers between a quarter and three-quarters of the height of the parent plant. The petiole base leaf blotches are small, brown, with a straight canal with erect margins. The petiole margin width is 1.5–2.0 cm, and the petiole is relatively short (44–55.50 cm).
The leaf blade length is 199–223.5 cm, the width is 58–81.5 cm, and the leaf ratio is 2.7. The shape of the leaf blade base is rounded on both sides. The upper surface of the leaf is shiny and very waxy. The lower surface of the leaf is yellow-green, with a dull appearance. The leaf midrib ventral and dorsal surfaces are yellow-green, with symmetric leaf blade insertion sites on the petiole.
The peduncle is medium length (39 cm), 3.8 cm in width, yellow-green, and hairless. The bunch hangs at a slight angle; it has a cylindrical shape and a very compact appearance. The male bud shape is ovoid. The male flower is yellow-orange. The male bud length at harvest is 27–29 cm. Bract base shape: tiny shoulder, obtuse apex, dark red exterior face, dark red interior face, deep grooves, and a waxy texture. Before falling, the bract rolls and creates very prominent scars on the rachis. Male flowers fall after the bracts. Free tepal is oval in shape, translucent white, obtuse, and has a developed apex. There are five anthers, which are yellow with red at the lobe margin, anther exertion is at the same level, and the filament is yellow-green. The ovary shape is curved, the basic color is yellow, the style is straight, and the stigma is yellow. The arrangement of the ovules is four-rowed.
A hand contains 15–18 fruits with a 10 cm fruit length. The shape is straight (or slightly curved). The transverse section of the fruit is round. The fruit apex is bottle-necked. The persistent style is the remains of the flower relicts at the fruit apex. The fruit pedicle length is 20 mm, the fruit pedicle width is 10 mm, and the pedicle surface is hairless. The color of the immature fruit peel is bright yellow-green. The mature fruit peel color is yellow. The fruit peel thickness is 2 mm. The pulp color before maturity is pale yellow. The flesh texture is firm. The predominant taste is astringent. The pulp color at maturity is yellow. The flesh texture is soft. The mature pulp is seedless, and the dominant taste is sugary.
The tallest plants with the largest leaves were observed in clone A. Height (368.5 ± 9.2 cm), pseudostem circumference (68.5 ± 3.5 cm), petiole length (51.5 ± 0.7 cm.), leaf blade length (223.5 ± 14.9 cm), and leaf blade width (81.5 ± 2.1 cm) are presented in Table 5. When six quantitative characteristics were evaluated, only the leaf blade width was statistically significant (p ≤ 0.05) (Table 5). Two agronomic characteristics were observed: the number of hands per bunch and the weight per bunch. Clone I had the highest number of hands (12.0 ± 2.03) per bunch and the greatest weight per bunch (17.0 ± 2.8 kg) followed by clones D, A, and B.

3.2. Genetic Variation Using Molecular Markers

3.2.1. ISSR-PCR

The except for primer E exhibited a monomorphic pattern, and the other primers (UBC-807, UBC-814, UBC-818, UBC-824, UBC-857, C, and D) produced polymorphic banding patterns (Figure 3). A total of 76 alleles and 62 polymorphic bands were observed. The alleles ranged from 350 to 2500 bp, with an average of 9.5 alleles per primer and an 80% polymorphism rate. The primer UBC-818 produced 13 polymorphic bands with a polymorphic percentage of 100%. The PIC values varied from 0.2550 to 0.4990. Thirty-six samples of Musa (AA, BB, and ABB) were analyzed using primer UBC-814 (Table 6).

3.2.2. SRAP-PCR

Of the 64 SRAP primer pairs, seven revealed distinct DNA bands. The SRAP primer sets (Table 3) yielded an average of nine fragments per primer combination, resulting in 65 bands for detecting genetic diversity (Table 7). A total of 65 bands were assessed, of which 57 (or 87%) were polymorphic. Alleles ranged from 100 to 1300 bp (Figure 4), and PIC values ranged between 0.2822 and 0.5.

3.3. Cluster Analysis

The ISSR similarity coefficient varied from 0.11 to 1.00 in the 36 samples. The coefficient of similarity between samples AA (‘Kluai Hom Champa’) and BB (‘Kluai Tani Dum’) was 0.11, and that between ABB samples (‘Kluai Nam Wa Mali-Ong’ and ‘Kluai Nam Wa Kabkhao’) and AA (‘Kluai Hom Champa’) was 0.12. The ABB samples (‘Kluai Nam Wa Mali-Ong’ and ‘Kluai Nam Wa Kabkhao’) shared a high similarity coefficient (0.91–1.00) with the ABB ‘Nam Wa’ sample. The ISSR cluster analysis divided the 36 accessions into two distinct clusters based on the similarity coefficient (Table 8). Cluster I contained only Musa ‘Hom Champa’ (AA), whereas Cluster II was divisible into two subgroups. Subgroup A included only Musa ‘Hom Chan’ (AA); subgroup B contained all ABB samples, including ‘Nam Wa Looksaileung’, Kabkhoa, Ubon, Pakchong 50, Nuanchan, and Yak. Musa BB included ‘Kluai Tani Dum’ and ‘Kluai Tani Nakhon Si Thammarat’ (Figure 5). SRAP data for the 36 samples revealed similarity coefficients ranging from 0.06 to 1.00. Compared with the ‘Hom Champa’ and ‘Kluai Nam Wa Mali-Ong’ samples, the other samples showed a lower similarity coefficient (0.06). Figure 6 shows that the dendrogram produced using SRAP markers contains two major clusters based on similarity coefficients (Table 9). The first cluster includes the ABB and BB genome accessions. The second cluster is entirely composed of AA accessions. SRAP markers clearly distinguished between the AA, BB, and ABB genomic groups and generated a higher percentage of polymorphism (87%) than ISSR markers (81.6%). However, SRAP and ISSR markers placed all the ABB accessions in the same cluster.

4. Discussion

Musa (ABB) ‘Mali-Ong’ is a triploid that belongs to the subgroup Pisang Awak. The fruits are used for dehydrated banana processing in the northern region of Thailand. The high demand for ‘Nam Wa Mali-Ong’ in northern Thailand exceeds supply. The cultivar has been replaced with other high-yielding cultivars from different parts of Thailand or neighboring countries. However, mixing Musa (ABB) cultivars is a significant problem in terms of quality for the dried banana industry. Seventy-five morphological characteristics were used to describe and characterize ‘Nam Wa Mali-Ong’. The general characteristics of ‘Nam Wa Mali-Ong’ are comparable to those of other ABB species, including ‘Nam Wa Looksaileung and Kabkhoa. ‘Nam Wa Mali-Ong’ has a thinner pseudostem, which has a height >3. The base of the leaf blade is rounded on both sides, with the upper surface being green and the lower surface yellow-green and extremely waxy. The peduncle is medium in length, yellow-green, and hairless. The ovule consists of four rows. Before maturation, the pulp is pale yellow; however, after maturation, it becomes yellow. These cultivars can be distinguished by the ripe pulp color; ‘Nam Wa Looksaileung’ is bright yellow, Kabkhoa is pale pink to white, and Mali Ong is yellow. The leaf blade width was the only quantitative attribute that was statistically distinct, and this distinction may have resulted from an epistatic or pleiotropic interaction [22]. The growth and yield of eight clones, including ‘Nam Wa Pak Chong 50’, ‘Nam Wa Mali Ong’, ‘Nam Wa Tanao Si’, ‘Nam Wa Ngoen’, ‘Nam Wa Khom’, ‘Nam Wa Thong Ma Eng’, and ‘Nam Wa Dam’, were documented [23]. The heights of the pseudostems of the eight clones ranged from 2.7 to 4.1 m. The pedicle length and peel thickness of distinct clones varied. Bunch weights ranged from 19 to 31 kg. Fruit was harvested 58–64 weeks after the planting date. ‘Nam Wa Mali-Ong’ was reported as having low bunch weights [23]. In this study, the first crop cycle of ‘Nam Wa Mali Ong’ yielded bunch weights of 12.17 to 17.00 kg. Bunch weight yield often increases between the second and third crop cycles [24]. The breadth of the leaf blades was observed to vary substantially. Clone D had the shortest leaf blade size; however, this did not affect the yield per bunch. This investigation clarified 77 morphological ‘Nam Wa Mali Ong’ characteristics.
The ISSR and SRAP markers showed high PIC values among the Musa accessions, ranging from 0.2550 to 0.4998 (average, 0.3850; Table 4) and from 0.2822 to 0.5 (average, 0.4527; Table 5), respectively. The PIC values indicated that ISSR and SRAP markers effectively revealed genetic variation among the banana cultivars. Compared with AFLP markers, SRAP markers provide three times more specific and unique bands [25]. According to a previous study, Musa (ABB) showed low variation when the genetic relationship was assessed using SSR [2]. SRAP markers are recommended as a more efficient means of distinguishing between Musa schizocarpa, M. balbisiana, and M. acuminata in the Eumusa section and between plantains and cooking bananas among triploid cultivars [26]. Previous research using ISSR and SCoT markers to investigate the genetic diversity of Musa species with different genomes showed that ISSR markers produced lower levels of genetic polymorphism [27]. Phitsanulok Province is in the low northern region of Thailand, comprising nine districts, Muang, Bang Rakam, Bang Kratum, Nakhon Thai, Chat Trakarn, Wang Thong, Neon-Maprang, Wat Bot, and Prom Phiram. Plains and plateaus dominate the geography of Phitsanulok Province. The highest point, Chart Trakan, is 800 m above sea level. The genetic variation of the accessions was not affected by the environment. Unlike morphological markers, molecular markers are unaffected by environmental factors [28]. The morphological characteristics and flowering of the ABB, BB, and AA groups require 12–16 months to be observed, and bunching occurs yearly. Environmental parameters regularly play a strong role in morphological expressions in plants, which occasionally may not correlate with molecular markers because of modifications in non-coding sequences. The leaf blade width separated the nine clones into four groups. The clones A, B, D, and I can be developed to produce an elite high yield cultivar that can be used in the processed food industry in the northern region of Thailand. Substitution of other high bunch yield ABB cultivars for ‘Nam Wa Mali-Ong’ may lead to its disappearance from fields. The findings of this study support that ex situ germplasm conservation in the northern region of Thailand should be urgently conducted on-farm and in vitro for the sustainable use of ‘Nam Wa Mali Ong’ in the processed food industry in Thailand. Novel Mali-Ong plants that are tolerant to diseases such as yellow sigatoka, anthracnose, and brown spot, which are destroying banana farms and reducing bunch yields, also need to be developed in the future. ISSR and SRAP markers effectively classified Musa AA, BB, and ABB cultivars. The data from the SRAP and ISSR markers and morphological characteristics produced corresponding results regarding the variation in Nam Wa Mali Ong.

5. Conclusions

This study described 77 morphological traits of Musa (ABB) ‘Mali-Ong’. Clones A, B, D, and I appeared to be superior with the highest bunch weight among the collected clones. SRAP markers were used to detect DNA variation in Musa AA, ABB, and BB groups. The ABB accessions were found to have a higher degree of genetic similarity and were clustered together. SRAP and ISSR makers can be employed to conserve and breed bananas.

Author Contributions

Conceptualization, D.P. and S.P.; methodology, T.B., A.K., K.S., and K.R.; software, T.B.; validation, D.P. and S.P.; formal analysis, D.P. and S.P.; investigation, D.P. and S.P.; resources, D.P. and S.P.; data curation, D.P. and S.P.; writing—original draft preparation, D.P. and S.P.; writing—review and editing, D.P. and S.P.; visualization, T.B., D.P. and S.P.; supervision, D.P. and S.P.; project administration, D.P.; funding acquisition, D.P. and S.P. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by Thailand Science Research and Innovation (TSRI) in the fiscal year 2021 (No. FRB 640025) and Naresuan University (Project code: R2564B003).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Acknowledgments

We thank the director of Plant Propagation Center No. 6 in Phitsanulok Province for providing a planting plot for bananas and to Wiramon Prangtong for coordinating with banana growers. Further appreciation is expressed to those who contributed to the banana collection.

Conflicts of Interest

The authors declare no conflict of interest.

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Figure 1. Twelve collection sites of Musa (ABB) ‘Mali-Ong’ across nine districts of Phitsanulok Province, Thailand. A: Mueang; B: Wang Thong; C: Bang Rakam; D: Wat Bot; E: Bang Krathum; F: Phrom Phiram; G: Noen Maprange; H: Nakhon Thai; and I: Chat Trakan.
Figure 1. Twelve collection sites of Musa (ABB) ‘Mali-Ong’ across nine districts of Phitsanulok Province, Thailand. A: Mueang; B: Wang Thong; C: Bang Rakam; D: Wat Bot; E: Bang Krathum; F: Phrom Phiram; G: Noen Maprange; H: Nakhon Thai; and I: Chat Trakan.
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Figure 2. Morphological characteristics of Musa (ABB) ‘Mali-Ong’. (A) Banana plant; (B) pseudostem color; (C) leaf blotches at petiole base; (D) leaf; (E) petiole canal leaf III; (F) color of bract external face; (G) color of bract internal face; (H) male bud; (I) male flower; (J) compound tepal color; (K) free tepal; (L) inflorescence; (M) bunch shape; (N) hand before maturity; (O) hand at maturity; (P) fruit at maturity; (Q) color of pulp before maturity; and (R) color of pulp at maturity.
Figure 2. Morphological characteristics of Musa (ABB) ‘Mali-Ong’. (A) Banana plant; (B) pseudostem color; (C) leaf blotches at petiole base; (D) leaf; (E) petiole canal leaf III; (F) color of bract external face; (G) color of bract internal face; (H) male bud; (I) male flower; (J) compound tepal color; (K) free tepal; (L) inflorescence; (M) bunch shape; (N) hand before maturity; (O) hand at maturity; (P) fruit at maturity; (Q) color of pulp before maturity; and (R) color of pulp at maturity.
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Figure 3. Amplification profiles of 36 banana samples using eight ISSR primers (UBC807, UBC814, UBC818, UBC824, UBC857, C, D, and E). M = 100 bp DNA ladder. Sample order (1–36 from left to right): 1= HCP; 2 = HC; 3 = TD; 4 = TNRT; 5 = NL; 6 = NU; 7 = NKK; 8 = NNJ; 9 = NPC50; 10 = NY; 11 = A021P01; 12 = A021P08; 13 = A011P08; 14 = A011P08; 15 = B091P04; 16 = B091P04; 17 = C041P03; 18 = C041P06; 19 = C051P06; 20 = C061P09; 21 = D071P03; 22 = D071P07; 23 = D081P05; 24 = D081P08; 25 = E031P01; 26 = 031P03; 27 = F061P03; 28 = F061P08; 29 = G121P03; 30 = G121P05; 31 = H101P03; 32 = H101P07; 33 = I11P04; 34 = I 111P08; 35 = P131P03; 36 = P131.
Figure 3. Amplification profiles of 36 banana samples using eight ISSR primers (UBC807, UBC814, UBC818, UBC824, UBC857, C, D, and E). M = 100 bp DNA ladder. Sample order (1–36 from left to right): 1= HCP; 2 = HC; 3 = TD; 4 = TNRT; 5 = NL; 6 = NU; 7 = NKK; 8 = NNJ; 9 = NPC50; 10 = NY; 11 = A021P01; 12 = A021P08; 13 = A011P08; 14 = A011P08; 15 = B091P04; 16 = B091P04; 17 = C041P03; 18 = C041P06; 19 = C051P06; 20 = C061P09; 21 = D071P03; 22 = D071P07; 23 = D081P05; 24 = D081P08; 25 = E031P01; 26 = 031P03; 27 = F061P03; 28 = F061P08; 29 = G121P03; 30 = G121P05; 31 = H101P03; 32 = H101P07; 33 = I11P04; 34 = I 111P08; 35 = P131P03; 36 = P131.
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Figure 4. Amplification profiles of 36 banana samples using seven pairs of SRAP primers (Me1 + Em6, Me1 + Em7, Me3 + Em5, Me6 + Em2, Me6 + Em3, Me6 + Em4, and Me9 + Em7). M = 100 bp DNA ladder. Sample order (1–36 from left to right): 1 = HCP; 2 = HC; 3 = TD; 4 = TNRT; 5 = NL; 6 = NU; 7 = NKK; 8 = NNJ; 9 = NPC50; 10 = NY; 11 = A021P01; 12 = A021P08; 13 = A011P08; 14 = A011P08; 15 = B091P04; 16 = B091P04; 17 = C041P03; 18 = C041P06; 19 = C051P06; 20 = C061P09; 21 = D071P03; 22 = D071P07; 23 = D081P05; 24 = D081P08; 25 = E031P01; 26 = 031P03; 27 = F061P03; 28 = F061P08; 29 = G121P03; 30 = G121P05; 31 = H101P03; 32 = H101P07; 33 = I11P04; 34 = I 111P08; 35 = P131P03; 36 = P131P.
Figure 4. Amplification profiles of 36 banana samples using seven pairs of SRAP primers (Me1 + Em6, Me1 + Em7, Me3 + Em5, Me6 + Em2, Me6 + Em3, Me6 + Em4, and Me9 + Em7). M = 100 bp DNA ladder. Sample order (1–36 from left to right): 1 = HCP; 2 = HC; 3 = TD; 4 = TNRT; 5 = NL; 6 = NU; 7 = NKK; 8 = NNJ; 9 = NPC50; 10 = NY; 11 = A021P01; 12 = A021P08; 13 = A011P08; 14 = A011P08; 15 = B091P04; 16 = B091P04; 17 = C041P03; 18 = C041P06; 19 = C051P06; 20 = C061P09; 21 = D071P03; 22 = D071P07; 23 = D081P05; 24 = D081P08; 25 = E031P01; 26 = 031P03; 27 = F061P03; 28 = F061P08; 29 = G121P03; 30 = G121P05; 31 = H101P03; 32 = H101P07; 33 = I11P04; 34 = I 111P08; 35 = P131P03; 36 = P131P.
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Figure 5. Dendrogram constructed using the unweighted pair-group method with arithmetic average (UPGMA) analysis based on ISSR similarity coefficients of 36 Musa samples.
Figure 5. Dendrogram constructed using the unweighted pair-group method with arithmetic average (UPGMA) analysis based on ISSR similarity coefficients of 36 Musa samples.
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Figure 6. Dendrogram constructed using UPGMA analysis based on SRAP similarity coefficients of 36 Musa samples.
Figure 6. Dendrogram constructed using UPGMA analysis based on SRAP similarity coefficients of 36 Musa samples.
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Table 1. List of Musa (ABB) ‘Mali-Ong’ samples collected from Phitsanulok Province, Thailand, and reference genomes AA and BB.
Table 1. List of Musa (ABB) ‘Mali-Ong’ samples collected from Phitsanulok Province, Thailand, and reference genomes AA and BB.
No.Local NameGenomic GroupCollection SiteCode
1.‘Kluai Hom Champa’AAPhitsanulokHCP
2.‘Kluai Hom Chan’AASukhothai Horticultural Research CenterHC
3.‘Kluai Tani Dam’BBPhitsanulokTD
4.‘Kluai Tani Nakhon Si Thammarat’BBSukhothai Horticultural Research CenterTNRT
5.‘Kluai Nam Wa Looksileuang’ABBPhitsanulokNYL
6.‘Kluai Nam Wa UbonABBPhitsanulokNU
7.‘Kluai Nam Wa KabkhaoABBPhitsanulokNKK
8.‘Kluai Nam Wa NuanchanABBPhitsanulokNNJ
9.‘Kluai Nam Wa Pakchong 50ABBResearch Center (Pakchong)NPC 50
10.‘Kluai Nam Wa YakABBResearch Center (Pakchong)NY
11.‘Kluai Nam Wa Mali-Ong’ ABBMueangA021P01
12.‘Kluai Nam Wa Mali-Ong’ ABBMueangA021P08
13.‘Kluai Nam Wa Mali-Ong’ ABBMueangA011P04
14.‘Kluai Nam Wa Mali-Ong’ ABBMueangA011P08
15.‘Kluai Nam Wa Mali-Ong’ ABBWang ThongB091P04
16.‘Kluai Nam Wa Mali-Ong’ ABBWang ThongB091P08
17.‘Kluai Nam Wa Mali-Ong’ ABBBang RakamC041P03
18.‘Kluai Nam Wa Mali-Ong’ ABBBang RakamC041P06
19.‘Kluai Nam Wa Mali-Ong’ ABBBang RakamC051P06
20.‘Kluai Nam Wa Mali-Ong’ ABBBang RakamC061P09
21.‘Kluai Nam Wa Mali-Ong’ ABBWat BotD071P03
22.‘Kluai Nam Wa Mali-Ong’ ABBWat BotD071P07
23.‘Kluai Nam Wa Mali-Ong’ ABBWat BotD081P05
24.‘Kluai Nam Wa Mali-Ong’ ABBWat BotD081P08
25.‘Kluai Nam Wa Mali-Ong’ ABBBang KrathumE031P01
26.‘Kluai Nam Wa Mali-Ong’ ABBBang KrathumE031P03
27.‘Kluai Nam Wa Mali-Ong’ ABBPhrom PhiramF061P03
28.‘Kluai Nam Wa Mali-Ong’ ABBPhrom PhiramF061P08
29.‘Kluai Nam Wa Mali-Ong’ ABBNoen MaprangG121P03
30.‘Kluai Nam Wa Mali-Ong’ ABBNoen MaprangG121P05
31.‘Kluai Nam Wa Mali-Ong’ ABBNakhon ThaiH101P03
32.‘Kluai Nam Wa Mali-Ong’ ABBNakhon ThaiH101P07
33.‘Kluai Nam Wa Mali-Ong’ ABBChat TrakanI111P04
34.‘Kluai Nam Wa Mali-Ong’ ABBChat TrakanI111P08
35.‘Kluai Nam Wa Mali-Ong’ ABBPak Chong,
Nakhon Ratchasima
P131P03
36.‘Kluai Nam Wa Mali-Ong’ ABBPak Chong,
Nakhon Ratchasima
P131P06
Table 2. Inter simple sequence repeat (ISSR) primers showing strong amplification and good reproducibility selected in this study.
Table 2. Inter simple sequence repeat (ISSR) primers showing strong amplification and good reproducibility selected in this study.
Primer NamePrimer Nucleotide Sequence
(5′-3′)
GCTmSource
UBC-807AGAGAGAGAGAGAGAGT4752[11]
UBC-814CTCTCTCTCTCTCTCTA4750[11]
UBC-818CACACACACACACACAG5350[11]
UBC-824TCTCTCTCTCTCTCTCG5350[15]
UBC-857ACACACACACACACACVG5050[16]
CGAGGAGGAGGAGGAGAC6552[17,18,19]
DGAGGAGGAGGAGGAGAT5950[17,18,19]
EGGGGTGGGGTGGGGT8050[18,19]
Table 3. Forward and reverse sequence-related amplified polymorphism (SRAP) primers and seven primer combinations applied in this study.
Table 3. Forward and reverse sequence-related amplified polymorphism (SRAP) primers and seven primer combinations applied in this study.
Primer Information
Forward primer
Me 1TGAGTCCAAACCGGATA
Me3TGAGTCCAAACCGGAAT
Me6TGAGTCCAAACCGGTAG
Me9TGAGTCCAAACCGGTCA
Reverse primer
Em2GACTGCGTACGAATTTGC
Em3GACTGCGTACGAATTGAC
Em4GACTGCGTACGAATTTGA
Em6GACTGCGTACGAATTGCA
Em7GACTGCGTACGAATTATG
Primer combination
Me1 + Em6
Me1 + Em7
Me3 + Em5
Me6 + Em2
Me6 + Em3
Me6 + Em4
Me9 + Em7
Table 4. Morphological characteristics of Musa (ABB) ‘Mali-Ong’. The evaluation was performed in Phitsanulok Province, Thailand using banana descriptors [14]. Color was determined using a Royal Horticultural Society (RHS) color chart.
Table 4. Morphological characteristics of Musa (ABB) ‘Mali-Ong’. The evaluation was performed in Phitsanulok Province, Thailand using banana descriptors [14]. Color was determined using a Royal Horticultural Society (RHS) color chart.
No.Morphological CharacteristicsDescription
1Leaf habitIntermediate, with normal leaves
2–6PseudostemAverage height, 326.77 ± 22.45 cm; average pseudostem circumference, 60.72 ± 6.7 cm; slender; color, yellow-green group 144 C; waxy
7–9Number of suckersEight suckers, close to parent, between ¼ and ¾ of the height of the parent plant
10–11Leaf blotches at petiole base Small blotches, color of blotches; brown group 200 A
12Petiole canal leaf IIIStraight with erect margins
13Petiole margin colorBrown group 200 A
14Petiole margin width1.5–2 cm
15Petiole length44–55.50 cm (short)
16Leaf blade length199–223.5 cm
17Leaf blade width58.5–81.5 cm
18Color of leaf upper surface Green group 137 B
19Appearance of leaf upper surfaceShiny
20Color of leaf lower surfaceYellow-green group 148B
21Appearance of leaf lower surfaceDull
22Wax on leavesVery waxy
23Insertion points of leaf blades on petiole Symmetric
24Shape of leaf blade base Both sides rounded
25Color of midrib ventral surfaceYellow-green group N144D
26Color of midrib dorsal surfaceYellow-green group 154 D
27–29Inflorescence/male budMedium peduncle length (39 cm), peduncle width 3.8 cm, peduncle color strong yellow–green 144D
30Peduncle hairinessPeduncle hairless
31Bunch positionSlightly angle
32Bunch shape Cylindrical
33Bunch appearanceVery compact
34Male bud size Length of male bud at harvest, 27–29 cm
35Bract base shapeSmall shoulder
36Bract apex shapeObtuse
37Color of bract external faceDark red, greyed-purple group 183 A
38Color of bract internal faceDark red, greyed-purple group 183 B
39Bract scars on rachisVery prominent
40Bract behavior before fallingRolling
41Wax on bractVery waxy
42Presence of grooves on the bractStrong grooved
43Male flower/male flower behaviorFalling after the bract
44Compound tepal basic colorRed-purple 60A
45Lobe color of compound tepalYellow 13 A
46Free tepal colorTranslucent white
47Free tepal shapeOval
48Free tepal apex developmentDeveloped
49Free tepal apex shapeObtuse
50Anther exertionIn relation to the base of the lobes on the compound tepal same level
51Filament colorYellow-green 11 D
52Anther colorYellow group 11 B with Red 39 B at margin
53Style basic colorYellow group 11 D
54Style shapeStraight
55Color of stigmaYellow group 11 B
56Ovary shapeCurved
57Ovary basic colorYellow group 11 C
58Arrangement of ovulesFour-rowed
59Fruit/number of Fruits 15–18
60Fruit length 10 cm
61Fruit shape Straight (or slightly curved)
62Transverse section of fruitRound
63Fruit apex Bottle-necked
64Remains of flower relicts at fruit apexPersistent style
65Fruit pedicle length20 mm
66Fruit pedicle width10 mm
67Pedicle surfaceHairless
68Immature fruit peel colorStrong yellow-green B
69Mature fruit peel colorYellow group 9 B
70Fruit peel thickness2 mm
71Pulp color before maturityYellow group 11D
72Flesh textureFirm
73Predominant taste Astringent
74Pulp color at maturityYellow group 8D
75Flesh textureSoft
76Predominant taste Sugary
77Presence of seed with source of
pollen
Seedless
Table 5. Quantitative characteristics of Musa (ABB) ‘Mali-Ong’ collected from nine farms.
Table 5. Quantitative characteristics of Musa (ABB) ‘Mali-Ong’ collected from nine farms.
CloneMorphological Characteristic
PseudostemPetioleLeaf Blade
Height
(cm)
Circumference
(cm)
Margin Width (cm)Length (cm)Length (cm)Width
(cm)
No. of Hand/BunchWeight/Bunch (kg)
A368.5 ± 9.268.5 ± 3.52.0 ± 0.051.5 ± 0.7223.5 ± 14.981.5 ± 2.1 a10.67 ± 0.0 abc14.67 ± 1.4 ab
B362.5 ± 17.767.5 ± 2.12.0 ± 0.049.5 ± 0.7199 ± 2.880.0 ± 1.4 ab9.0 ± 0.7 abc13.67 ± 0.7 ab
C323.5 ± 26.249.0 ± 2.81.5 ± 0.744.0 ± 4.2175.5 ± 19.158.5 ± 5.7 d8.0 ± 0.0 c12.17 ± 0.4 b
D311.5 ± 26.259.5 ± 3.51.75 ± 0.455.5 ± 2.1183.5 ± 29.059.0 ± 5.7 d11.7 ± 0.7 ab15.7 ± 0.7 ab
E320.0 ± 28.363.5 ± 3.52.15 ± 0.247.5 ± 6.4213.5 ± 10.674.5 ± 3.5 abc8.67 ± 1.4 bc12± 2.8 b
F282.5 ± 53,053.0 ± 12.72.0 ± 0.055.5 ± 6.4181 ± 33.966.8 ± 8.1 cd8 ± 0.0 c12.2 ± 0.0 b
G315.0 ± 26.956.25 ± 3.91.75 ± 0.451.5 ± 2.1202.5 ± 27.664.8 ± 7.4 cd8.7 ± 1.4 bc12.7 ± 1.4 b
H314.0 ± 2.861.5 ± 0.71.8 ± 0.454.5 ± 10.6199.5 ± 0.769.5 ± 0.7 abc8.0 ± 0.0 c12.2 ± 0.0 b
I343.5 ± 12.067.8 ± 2.52.0 ± 0.049.5 ± 0.7212.5 ± 20.572.8 ± 1.1 abc12.0 ± 2.0 a17.0 ± 2.8 a
nsnsnsnsns***
Data are mean values ± standard deviation of three biological replicate of each clone. Lowercase letters are valid for the same column. ANOVA was used to test for significance, and significant differences between means were tested by DMRT and are indicated by different letters (p ≤ 0.05). ns = non-significant and * = significant.
Table 6. Polymorphism obtained from eight ISSR primers among 36 Musa samples.
Table 6. Polymorphism obtained from eight ISSR primers among 36 Musa samples.
No.ISSR
Primer
Allele Size Range (bp)Amplified BandsPIC
Value
TotalPolymorphicPolymorphism (%)
1.UBC-807450–150011981.810.4662
2.UBC-814550–25009777.780.4998
3.UBC-818350–180013131000.3200
4.UBC-824400–2300991000.4968
5.UBC-857350–2300108800.4278
6.C550–1700106600.3318
7.D450–16007342.860.2550
8.E650–1700771000.2822
Total 7662 3.0796
Average 9.57.7580.310.3850
PIC—polymorphic information content.
Table 7. Polymorphism obtained from seven SRAP primer combinations among 36 Musa samples.
Table 7. Polymorphism obtained from seven SRAP primer combinations among 36 Musa samples.
No.SRAP PrimerAllele Size Range (bp)Amplified BandsPIC Value
TotalPolymorphicPolymorphism (%)
1.Me1 + Em6150–1100111090.90.4992
2.Me1 + Em7100–12009888.880.4928
3.Me3 + Em5150–1300121191.660.4838
4.Me6 + Em2200–1300881000.2822
5.Me6 + Em3250–1300881000.4352
6.Me6 + Em4100–1300108800.4758
7.Me9 + Em7150–8007457.140.5000
Total 6557 3.1690
Average 9.298.1486.940.4527
Table 8. Similarity coefficient matrix of 36 Musa samples based on ISSR primers.
Table 8. Similarity coefficient matrix of 36 Musa samples based on ISSR primers.
HJPHCHTDTNRTNLYNUNKKNNJNPC50NVYA11A12A21A22B11B12C11C12C21C22D11D12D21D22E11E12F11F12G11G12H11H12I11I12CT1CT2
HJP1.00
HCH0.291.00
TD0.110.331.00
TNRT0.100.350.971.00
NLY0.100.280.600.621.00
NU0.140.330.630.640.911.00
NKK0.120.300.580.600.970.941.00
NNJ0.140.330.630.640.911.000.941.00
NPC500.120.310.650.670.870.970.900.971.00
NVY0.140.330.630.640.911.000.941.000.971.00
A110.120.300.580.600.970.941.000.940.900.941.00
A120.120.300.580.600.970.941.000.940.900.941.001.00
A210.120.300.580.600.970.941.000.940.900.941.001.001.00
A220.120.300.580.600.970.941.000.940.900.941.001.001.001.00
B110.120.300.580.600.970.941.000.940.900.941.001.001.001.001.00
B120.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.00
C110.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.00
C120.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.00
C210.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.00
C220.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.00
D110.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.00
D120.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.00
D210.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.00
D220.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.00
E110.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.001.00
E120.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.00
F110.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.00
F120.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.00
G110.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.00
G120.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.00
H110.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.00
H120.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.00
I110.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.00
I120.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.00
CT10.130.280.600.620.930.910.970.910.870.910.970.970.970.970.970.970.970.970.970.970.970.970.970.970.970.970.970.970.970.970.970.970.970.971.00
CT20.120.300.580.600.970.941.000.940.900.941.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.001.000.971.00
Table 9. Similarity coefficient matrix of 36 Musa samples based on SRAP primer combination.
Table 9. Similarity coefficient matrix of 36 Musa samples based on SRAP primer combination.
HJPHCHTDTNRTNLYNUNKKNNJNPC50NVYA11A12A21A22B11B12C11C12C21C22D11D12D21D22E11E12F11F12G11G12H11H12I11I12CT1CT2
HJP1.00
HCH0.591.00
TD0.050.171.00
TNRT0.060.220.881.00
NLY0.140.290.590.541.00
NU0.120.260.690.630.881.00
NKK0.110.240.680.580.820.881.00
NNJ0.120.260.640.630.880.940.881.00
NPC500.130.280.670.660.850.850.790.851.00
NVY0.140.240.560.600.610.720.670.720.741.00
A110.100.220.700.600.850.850.910.850.820.701.00
A120.100.220.700.600.850.790.850.790.880.650.881.00
A210.090.210.730.630.830.880.940.880.800.680.910.911.00
A220.100.220.700.600.850.910.910.910.770.650.880.880.971.00
B110.100.220.700.600.850.910.910.910.820.700.940.880.910.941.00
B120.110.240.680.580.820.880.940.880.740.620.850.850.940.970.911.00
C110.090.240.680.630.880.880.820.880.790.670.910.850.880.910.910.881.00
C120.070.210.670.630.830.880.880.880.850.730.910.850.880.850.910.820.881.00
C210.090.210.680.630.820.880.880.880.850.780.910.850.880.850.910.820.880.941.00
C220.090.240.680.630.880.940.880.940.850.720.910.850.880.910.970.880.940.940.941.00
D110.080.220.700.650.850.910.910.910.880.750.940.880.910.880.940.850.910.970.970.971.00
D120.110.250.660.610.910.970.910.970.820.690.880.820.910.940.940.910.910.910.910.970.941.00
D210.120.260.640.580.940.820.820.820.910.660.910.910.820.790.850.770.880.880.880.880.910.851.00
D220.080.220.700.650.850.910.910.910.880.750.940.880.910.880.940.850.910.970.970.971.000.940.911.00
E110.090.240.680.630.820.880.880.880.850.780.910.850.880.850.910.820.880.940.940.940.970.910.880.971.00
E120.060.200.700.650.800.850.850.850.830.710.880.820.850.820.880.790.850.970.910.910.940.880.850.940.911.00
F110.070.210.730.680.830.880.880.880.850.730.910.850.880.850.910.820.880.940.940.940.970.910.880.970.940.971.00
F120.070.210.730.680.830.880.880.880.850.730.910.850.880.850.910.820.880.940.940.940.970.910.880.970.940.971.001.00
G110.080.220.700.650.850.850.850.850.820.700.940.820.850.820.880.790.910.910.910.910.940.880.910.940.910.940.970.971.00
G120.070.210.730.680.830.880.880.880.850.730.910.850.880.850.910.820.880.940.940.940.970.910.880.970.940.971.001.000.971.00
H110.110.250.660.610.910.970.910.970.820.690.880.820.910.940.940.910.910.910.910.970.941.000.850.940.910.880.910.910.880.911.00
H120.110.250.660.610.910.970.910.970.820.690.880.820.910.940.940.910.910.910.910.970.941.000.850.940.910.880.910.910.880.911.001.00
I110.090.240.680.630.880.940.880.940.790.670.850.850.940.970.910.940.940.880.880.940.910.970.820.910.880.850.880.880.850.880.970.971.00
I120.080.220.650.600.850.910.850.910.770.650.820.820.910.940.880.910.910.910.850.910.880.940.790.880.850.880.850.850.820.850.940.940.971.00
CT10.110.220.710.660.690.690.740.690.660.640.760.710.740.710.710.740.790.740.790.740.760.710.740.760.740.770.790.790.820.790.710.710.740.711.00
CT20.060.170.700.650.750.800.790.800.770.710.820.820.850.820.820.790.850.910.910.850.880.820.800.880.850.940.910.910.880.910.820.820.850.880.821.00
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Premjet, D.; Boonsrangsom, T.; Sujipuli, K.; Rattanasut, K.; Kongbungkerd, A.; Premjet, S. Morphological and Molecular Characterizations of Musa (ABB) ‘Mali-Ong’ in Thailand. Biology 2022, 11, 1429. https://doi.org/10.3390/biology11101429

AMA Style

Premjet D, Boonsrangsom T, Sujipuli K, Rattanasut K, Kongbungkerd A, Premjet S. Morphological and Molecular Characterizations of Musa (ABB) ‘Mali-Ong’ in Thailand. Biology. 2022; 11(10):1429. https://doi.org/10.3390/biology11101429

Chicago/Turabian Style

Premjet, Duangporn, Thanita Boonsrangsom, Kawee Sujipuli, Kumrop Rattanasut, Anupan Kongbungkerd, and Siripong Premjet. 2022. "Morphological and Molecular Characterizations of Musa (ABB) ‘Mali-Ong’ in Thailand" Biology 11, no. 10: 1429. https://doi.org/10.3390/biology11101429

APA Style

Premjet, D., Boonsrangsom, T., Sujipuli, K., Rattanasut, K., Kongbungkerd, A., & Premjet, S. (2022). Morphological and Molecular Characterizations of Musa (ABB) ‘Mali-Ong’ in Thailand. Biology, 11(10), 1429. https://doi.org/10.3390/biology11101429

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