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Lansium Parasiticum (Osbeck) K.C

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Lansium Parasiticum (Osbeck) K.C RESEARCH ARTICLE SABRAO Journal of Breeding and Genetics 51 (2) 151-160, 2019 THE ABSOLUTE GENETIC INVARIABILITY OF POLYEMBRYONY SEEDLING OF DUKU ‘KUMPEH’ (Lansium parasiticum (Osbeck) K.C. Sahni & Bennet.), A SUPERIOR TYPE FROM JAMBI, INDONESIA P. MURNI1, SYAMSUARDI2*, NURAINAS2, E. MANSYAH³ and M. HUDA² 1Department of Biology, Faculty of Education, Universitas Jambi, Indonesia 2Departement of Biology, Faculty of Mathematics & Natural Sciences, Universitas Andalas, Indonesia 3Indonesian Tropical Fruit Research Institute, Solok, West Sumatra, Indonesia *Corresponding author’s email: [email protected] Email addresses of coauthors: [email protected], [email protected], [email protected], [email protected] SUMMARY Duku ‘Kumpeh’ is one of the superior and potential cultivar of Duku (Lansium parasiticum) that originate from a specific area in the Jambi and it becomes one of the essential sources for income of Jambi people. For sustainable utilization of this essential local germplasm of tropical fruit need the understanding of the phenomenon of polyembryony and genetic information of the plant. The study of genetic variation of multiple seedlings of duku from Kumpeh was conducted. In this study, 34 seedlings were used (14 seedlings from Kumpeh and 20 seedlings from Dusun Tuo) from two mother trees in order to collect the samples of DNA. Extracted DNA samples used a modification of CTAB method then amplified using four primers of Inter-Simple Sequence Repeat (ISSR) namely (AAC)6, (AAG)6, (AAT)10, (AG)10 and two from RAPD markers (OPA 13 and OPA 18) to asses genetic variation. Base on observed and analysis of genetic variation of samples showed that multiple seedlings of seven seeds (14 seedling individuals) from Kumpeh and ten seeds (14 seedling individuals from Dusun Tuo) have the uniform of DNA band profile and the same as those of mother trees. The existing identical of all seedlings suggested that polyembryony of duku seeds derived from the non-zygotic embryo. Thus, we concluded that the reproduction system of duku (Lansium parasiticum (Osbeck) K.C. Sahni and Bennet.) occurred asexual with apomictic reproduction. Key words: Lansium parasiticum, genetic uniformity, polyembryony, non-zygotic embryo Key findings: The polyembryony of duku was a potential mechanism to produce a high number of seedlings with good quality and their genetic uniformity. The existing identical of all seedlings from polyembryony suggested the embryo derived from the non-zygotic embryo. The reproduction system of duku (Lansium 151 SABRAO J. Breed. Genet. 51 (2) 151-160 parasiticum (Osbeck) K.C. Sahni and Bennet.) was asexual reproduction (apomictic). Manuscript received: February 24, 2019; Decision on manuscript: March 7, 2019; Accepted: March 30, 2019. © Society for the Advancement of Breeding Research in Asia and Oceania (SABRAO) 2019 Communicating Editor: Prof. Naqib Ullah Khan INTRODUCTION (Kishore, 2014). About 255 genera belonging to 153 families of Duku (Lansium parasiticum) is a Angiosperm plants were reported to tropical fruit tree species belongs to exhibit the polyembryony (Carman, the family of Meliaceae. The 1997). However, the seedlings from distribution of this species in South- polyembryony of each plant species East Asia mainly Peninsular Thailand, indicated the genetic variability Malesia, Indonesia especially Sumatra between them. and Java, Borneo, and Malay Generally, the seedlings of duku Peninsula (Mabberley et al., 1995), resulted from germination, and 25% but this species was distributed until of those seedlings resulted from Kuba, Honduras, India, Puerto Rico, polyembryony (Salma and Razali, Suriname, Thailand, Trinidad, Tobago, 1987). So it means the utilization of Amerika and Vietnam (Orwa et al., polyembryony to produce seedlings of 2009). duku still limited and application Duku ‘Kumpeh’ is one of the polyembryony to obtain the more superior and potential cultivars of seedlings is prospective for duku Duku (Lansium parasiticum (Osbeck) plantation. However, the seedlings K.C. Sahni and Bennet.) that originate resulted from polyembryony indicate from a specific area in the Jambi, genetic variability due to the zygotic namely Kumpeh village and it become process. In order to produce a good one of an essential source for income quality of this tropical fruit, the of Jambi people. Until now, commonly understanding of genetic uniformity is the local people have still produce the fundamental. seedlings from the seeds that why the Some seedlings from time for harvesting was longer. polyembryony were resulted from However, some cultivation from a zygotic and non-zygotic embryo vegetative organ such as cuttings, and process and will affect their genetic grafts were not a success due to the uniformity. For sustainable utilization problems of the thin of stem bark and of this essential local germplasm of the difficulty to remove from the tropical fruit, the information about wood. The seedling from genetic uniformity of seedlings from polyembryony of duku (Salma and polyembryonic was crucial. The Razali, 1987) was an alternative way purpose of this study was to detect to produce multiple seedlings from a the level of genetic uniformity of duku single seed during germination. seedlings as resulted from a specific Polyembryony plays a vital role in pattern of polyembryony of a potential horticulture and plant breeding type of duku from Kumpeh, Jambi. 152 Murni et al. (2019) MATERIALS AND METHODS the individual tree of Dusun Tuo were germinated in the greenhouse to This study used the samples of seeds obtain thirty-four seedlings of duku. collected from two mother trees in Based on their size, seedlings by Kumpeh (KP) and Dusun Tuo (TB) Kishore (2014) classified into two during a fruiting season in 2016 types. Initial individuals emergence (Figure 1). The position site is at S: with more significant big presumed as 01o34.254’- E: 103o51.350’ Kumpeh a zygotic seedling (hereafter and at S: 01o23.112’- E: 102o20.922’ mentioned by A-Type), in case of Dusun Tuo, Jambi province Indonesia Individuals the subsequent emergence (Murni et al., 2016). Totally seventeen with smaller in size presumed as non- seeds (seven seeds from the individual zygotic seedlings (B-Type). tree of Kumpeh and ten seeds from Figure 1. The collection site of two mother trees from Kumpeh and Dusun Tuo (Tebo), Jambi Province, Indonesia (Modification from http://www.google.com/ maps/place/Jambi, Indonesia). The young leaves of those (PVP). The DNA concentration was seedlings were extracted using CTAB measured by subjecting the samples (cetyl trimethyl ammonium bromide) into 1.2% agarose gel electrophoresis, method to obtain genomic DNA of staining with ethidium bromide and progenies (Doyle and Doyle, 1990). visualization on ultraviolet (UV) About 0.1 mg fresh young leaves was transilluminator used Bio-Doc Analyser ground for total DNA extraction by (BDA) Biometra. addition 1% of polyvinylpyrrolidone 153 SABRAO J. Breed. Genet. 51 (2) 151-160 The simple, rapid and multiple and compared between A and B type polymorphic loci ISSR and RAPD from different mother tree. The markers were applied in this study. seedlings were assumed as non- The four polymorphic ISSR primers zygotic embryo if the band pattern of [(AAC)6, (AAG)6, (AAT)10, and (AG)10] the progeny was no different to those (Murni et al., 2016) and two patterns of bands the mother tree. polymorphic RAPD primers, OPA-13 The differences pattern of the bands (5’CAG CAC CCA C3’) and OPA 18 (5’ will be analyzed and the number of AGG TCA CCG T 3’) (Yulita, 2011) bands per primer, the polymorphic were used in this study. PCR and monomorphic band was identified. amplification of DNA samples using The banding pattern between the ISSR and RAPD primers conducted seedling of progeny and mother tree with a total volume of 12.5 ml solution were compared. containing 6.25 mL of Taq Go Green The bands of progeny and Master Mix Promega, 4.25 mL of dd mother trees were scored, present of H2O (Nuclease-Free Water), 1 mL of band = 1 and no detected of band = primer, and 1 mL of sample DNA 0. The pattern of banded was (template), Amplification using PCR arranged as a biner data and analyzed thermocycler, GSX1 nexus Eppendorf to produce the degree of genetic Master cycler with 45 cycles. The variation using POPGENE version 1.32. reactions was initially with The genetic distance between progeny denaturation step at 95°C for one and the mother tree was calculated minute, a denaturation step at 95°C with the Jaccard similarity index (Yeh for one minute, annealing at et al., 1997). temperatures appropriate optimization for one minute, extension at 72°C for two minutes, and a final extension at RESULTS 72°C for 10 minutes. The results of PCR amplification subjected to Thirty-four seedling from seventeen electrophoresis on a 1.2% agarose gel seeds (seven seeds from Kumpeh and immersed in TBE buffer and running ten seeds from Tebo) produced one to with 110 V. Furthermore, applied six embryos. Based on the number of electrophoresis gel was stained with an embryo produced from two 1% Ethidium bromide for 10 minutes collection sites, the frequency number and washed by soaking in distilled of seedling were statistically different water for 15 minutes. Finally viewed between Kumpeh and Dusun Tuo under ultraviolet light and (Mann-Whitney P < 0.01). Duku from photographed using BDA Biometra. Kumpeh had a higher percentage of polyembryony (2-6 embryo) than Data analysis duku from Dusun Tuo (Figure 2a), the percentage of the monoembryo was The difference frequency of the found more in duku from Dusun Tuo. number of seedling per seed of Duku Based on this, the duku seeds from Kumpeh between the two collection Kumpeh more able to produce sites was statistically analyzed using polyembryony seeds. the Mann-Whitney U Test on the The observation of seedling degree of 95% confidence. The profile performance from germination of 17 of DNA from all seedlings observed seeds indicated that there were 154 Murni et al.
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