ATAVISM OBSERVED FROM MELON
(Cucumismelo L. cv. Hikadi)IN SELF-CROSS OF F1 HK.10 BY OPEN POLLINATION
IbnuSinaa, Budi S. Daryonoa,
aGenetics Laboratory, Faculty of Biology, Universitas Gadjah Mada,
Jl. Teknika Selatan, Sekip Utara, Daerah Istimewa Yogyakarta,55281, Indonesia
Corresponding e-mail:
Abstract
Melon (Cucumis melo L.) is one of the main seasonal fruit crops in Indonesia. Faculty of Biology Universitas Gadjah Mada had developed a new cultivated variety called Hikadi, result of further studies of Gama Melon perfume. Atavism is a process at genetic level that brings back either morphological or physiological traits from ancestor which had disappeared a few generations earlier. This research aimed to identify atavism occurrence in selfcross of F1 Hikadi cultivarsby open pollination grown in Gunungkidul, Yogyakarta, Indonesia. Analysis of phenotypic characters has been done by Unweighted Pair Group Method with Arithmetic Means (UPGMA) using similarity index (SSM) to the referenced Hikadi cultivar and its ancestor characters using melon’s characters list from the International Plant Genetic Resources Institute (IPGRI) and the Indonesia departement of agriculture’s Plant Variety Protection (PVP) as well as for quantitative phenotypic character analysis using chi-square test. Result shows that 4 out of 748 plants (0,53% of population) observed as atavism with 38,59% difference of similarity index (SSM) to the referenced Hikadi cultivar and significant difference in fruit’s weight. Phenotipic morphological characters that appeared shows similarity to one of its ancestor, Miyamauri-5 (MR-5).
Keywords: Melon; Hikadi; Atavism; Indonesia.
1. Introduction
Agriculture is one of the key sectors in Indonesian economy, especially the fruit and vegetable crops. Melon (CucumismeloL.) is one of the main seasonal fruit crops that included in Cucurbitaceae family. It’s been well-known for its juicy texture and sweet taste. 100 grams of melon contains 23,0 kcal, 0,6 g protein, 17 mg calcium, 2.400 IU vitamin A, 30 mg vitamin C, 0,045 mg thiamin, 0,065 mg ribloflavin, 1,0 mg niacin, 6,0 g carbohydrate, 0,4 mg ferrum, 0,5 mg nicotinamides (B12), 0,4 g fiber, and 93 g water (Tjahjadi, 1992).
Until this paper was written, there were only 79 melon cultivar that has been listed at Database of Plant Variety Protection made by Kementrian Pertanian Republik Indonesia (Deptan RI, 2015). Based on that fact, new melon cultivar development is needed to support melon farming especially in Indonesia.Biology Faculty of Universitas Gadjah Mada as centre of excellence in tropical biology further developed a new cultivated variety of melon called ‘Hikadi’, product of Gama Melon Parfum’sbreeding research and tested through multilocation and multiseasonal test.Selection to the favoured phenotipic charactersin plant ennoblement is a common thing to do. In this research, we used independent culling levelmethod to selects two or more characters at several generations consecutively.This method provides minimum level of specific phenotipic characters to be inherited through selective cross-breeding (Soemartono & Sardiyah,1998).
In assemblance of ‘Hikadi’ cultivar, the inherited fruit’s phenotipic characters expected to be relatively small (300-700 grams), crunchy and yellow-coloured fruit’s meat, low water contains, sweet, have mild aromatic character, and can endured long periode of storage (up to 14 days) at room temperature.
In assembling and breeding cultivars, it’s been well known there would be genetic instability in self-breeding products as a result of gene interaction from parental’s variety, or an epigenetic interference. One of the possible phenomenon that could happened is atavism. Atavism is a process in genenetic level that brought back either morphological, anatomical, or physiological characters that once belong to one of the parental and has been lost for several generations (Verhulst, 1996). Atavism in plant ennoblement is a phenomenon that could give a profit on its own, one of them is availability of parental’s character that probably has been lost for several generations before to be reused as prime characters source.
This research aim to study atavism case using morphological character of suspected F1‘Hikadi’ cultivar produced in self hybrid of HK.10F and HK.10C by sopen pollination which has been planted on Kemadang, Gunungkidul. This resesearch later hoped to be able to support Hikadi cultivar development, further research of genetic diversity enrichment, and melon plantennoblement as well as become reference to another melon cultivated variety development.
2. Material and Method
Research was conducted in October 2014 - January 2015 in Kemadang, Gunungkidul, Yogyakarta and Genetics Laboratory of the Faculty of Biology Universitas Gadjah Mada. The tools used in this study include bamboostands, raffia rope, camera, scales, trays, medline, ruler, shovels, buckets, stationery, logbook, and plastic to contain seeds. Materials used in this study is melon seed of HK.10C and HK.10F from Hikadi cultivars, water, fertilizer, fungicides, insecticides, mulches, and labels.
The first stage of this study is preparation; includes soil preparation before planting and germination of seeds. Land preparation is done by plowing so that the soil becomes loose and re-oxygenated. Furthermore, fertilization with manure land base (ZA, KCl and TS) had been done with a dose of ZA 1000 kg / ha, KCL and TS of 350 kg / ha. Provision of basic fertilizer was made during land preparation intended to provide sufficient nutrients for plants melons. Melon seed germination cultivars begins with incubating the seeds in a warm room on a tray by sand, cloth, water, and a 10 watt lamp for 24 hours. After that, sprouts transferred into a polybag seedlings until 10 days old and ready to be moved to be planted in the field.
The next stage of this research is the nurturing and maintenance of'Hikadi' melon plants. At the beginning of the planting process, the land is wateredand also carried out with the addition of dolomite to stabilize the pH of the soil. Each week there were two times of insecticides and fungicides sprays to prevent the provision of pests and diseases. Fertilizer application is given once in a week during week-old melon plants and after a 5-week-old melon plants, the dose plus 20% of the initial dose. After 60-65 days, there had been done selection of fruit above sections 9, 10, and 11. The melon fruits further ready to be harvested, sampled, and analyzed.
Sampling has been done by taking 10 pieces of fruit samples as the result of self-breeding F1 HK.10C & HK.10F randomly for morphological mean characters observation. After that, there wereselective samplingto the fruit of plants suspected of having atavism.
After sampling the suspected atavism then performed characterization consists of quantitative as well as qualitative characterization, which melons were observed and compared in accordance with the characters list of PVP (Plant Variety Protection) from Indonesia’s Agricultural Departement.
3. Result and Discussion
From the research that has been done, it is known there are 4 out of 748 plants (0.53% of the total population) suspected of having atavism (Figure 1).
Figure1. View from above (a), below (b), lateral (c) and transversal (d) of selected M1 fruit that indicates atavism. (Personal documentation, 2015)
When compared with the genealogical tree of ‘Hikadi’ melon, morphological fruit character results are of fruits that indicates atavism have common appereance with one of the main parent of the first generation, namely Miyamauri-5 (MR-5). It is characterized mainly by similiar qualitative phenotypic appearance,which is shown to forms a pattern of a vertical longitudinal lines of light green alternated by dark green lines, has no turbines, and has a lobe on the fruit (Figure 2).
Figure 2. Comparison between the selected M1 fruits that experiencing atavism (a) with elders from the first generation, Miyamauri-5 (b) (Personal Documentation, 2015; Daryono, 2013; Daryono & Supriyadi, 2012)
The comparison of qualitative morphological character of common Hikadi with selected fruits which indicates atavism shown in Table 1.
Table 1. Comparison of qualitative morphological character from common Hikadi cultivar with selected fruits which indicates atavism
No. / Character / M1 / M2 / M3 / M4 / HikadiOutside morphology
1 / Fruit’s shape / globular / globular / globular / globular / globular
2 / Younger skin’s color / Dark green / Dark green / Dark green / Dark green / Green-white
3 / Older skin’s color / Yellowish green / Yellowish green / Yellowish green / Yellowish green / Yellow-orange
4 / Net level / 0 / 1 / 0 / 5 / 1
5 / Color pattern / exist / exist / exist / exist / inexistent
6 / Skin’s surface pattern / seperti bola basket / seperti bola basket / seperti bola basket / seperti bola basket / polos dgn sdkt corak
7 / Vertical line / 10 / 10 / 12 / 10 / -
8 / Lobus availability / exist / exist / exist / exist / -
9 / Number of lobus / 10 / 10 / 12 / 10 / -
10 / Turbin availability / - / - / - / - / -
11 / Turbin’s shape / - / - / - / - / -
Inside morphology
12 / Seed cavity’s shape / Oval / oval / oval / pear / oval
13 / Fruit meat color / green / Yellowish green / green / Yellowish green / orange
14 / Fruit meat texture / crunchy / crunchy / crunchy / crunchy / Slightly juicy
15 / Water contains / low / low / low / low / medium
16 / Fruit meat taste / Vapid / Slightly sweet / Slightly sweet / Slightly sweet / sweet
17 / Fruit’s aroma / - / - / - / - / mild
18 / Seed color / white / white / white / white / white
As seen from Table 1 that there’s a distinctive qualitative morphological characters between common Hikadi with selected fruits which indicates atavism in color, pattern color, and skin surface. Furthermore, the data analyzed using calculation of the similarity value from atavistic fruit and common Hikadi.
The level of similarity thus determined by using Unweighted Pair Group Method with Arithmetic Means (UPGMA) by Simple Matching coëficient (Ssm) method. The formula used to calculate is as follows:
Ssm = ( a + d ) x 100%
( a + b + c + d )
(Bergey, 2001)
Explanation :
a = sum of characters that shown as (+) dan (+).
b = sum of characters that shown as (+) dan (-).
c = sum of characters that shown as (-) dan (+).
d = sum of characters that shown as (-) dan (-).
The value of each Ssm index similarity calculation incorporated into the similarity matrix clustering in the table followed by cluster analysis using average linkage clustering algorithm. Based on clustering analysis then the similarity of qualitative phenotypic characters representd by dendogram construction. Furthermore, to determine the feasibility of dendogram construction results, it’s tested by cophenetic correlation calculations in order to get the value of r. If the known value of r close to or equal to 100%, then the Ssm similiaritas index analysis which has been done indicates high level of confidence (reliability).
R value is obtained from the formula:
r = n ∑xy - ∑x.∑y x 100 %
√ {n ∑x² - (∑x)²}{n ∑y² - (∑y)²}
(Fisher & Yates, 1963).
Results of Ssm similarity index analysis for qualitative morphological characters from common Hikadi cultivar with selected fruits which indicates atavism shown in form of dendogram below (Figure 3) with r = 99.87%. This shows that the obtained dendogram shows high level of confidence (reliability).
Figure 3. A dendogram which shows the result of the analysis for qualitative morphological characters similarity
From dendogram that has been made, there were 36.59% difference of qualitative phenotypic characters from common Hikadi cultivar with selected fruits which indicates atavism. This shows the difference in the phenotype of fruit appereance is quite distant and conspicuous.
Afterwards, the quantitative phenotypic characters were examined. Comparison of quantitative phenotypic morphological characters from common Hikadi cultivar with selected fruits which indicates atavism shown in Table 2.
Table 2. Quantitative morphological character from selected fruits which indicates atavism
No / Characters / Unit / M1 / M2 / M3 / M4 / Mean / ±(stdev)1 / Weight / gram / 360 / 280 / 180 / 400 / 305,00 / 84,11
2 / Horizontal circumference / cm / 29 / 28 / 23,5 / 31 / 27,88 / 2,75
3 / Vertical circumference / cm / 30 / 28 / 24 / 30 / 28,00 / 2,45
4 / Top diameter / cm / 1 / 1 / 0,7 / 1 / 0,93 / 0,13
5 / Bottom diameter (Turbin) / cm / 2 / 1 / 1 / 1,3 / 1,33 / 0,41
6 / Horizontal length / cm / 9 / 8,5 / 7,5 / 10 / 8,75 / 0,90
7 / Vertical length / cm / 9,5 / 8,5 / 7,5 / 9,5 / 8,75 / 0,83
8 / Outer skin thickness / cm / 0,1 / 0,2 / 0,1 / 0,3 / 0,18 / 0,08
9 / Fruit meat’s thickness / cm / 2 / 2,1 / 1,7 / 2,7 / 2,13 / 0,36
From a quantitative calculation results which have been obtained, the data further analyzed using the chi-square test to having an understanding whether the character of common melon Hikadi cultivars with samples that undergo atavism has real difference or not (Table 3).
Chi-square test was a method widely used to test whether a set of data follows the Gaussian distribution or not. Chi square value is determined by the following equation:
(Fisher & Yates, 1963)
With Xi is the observed value of each measurement and X is the mean value of the expected measurement. The value of chi-square (x2) of the above calculation then matched to the chi-square table. If only the obtained value has exceed certain numbers of P-value at certain reliability (in this case, we use 95% level of confidence) from the table, it’s then decided the difference of quantitative characters observed was significant.
Tabel 3.Comparison of mean quantitative morphological character from common Hikadi cultivar with selected fruits which indicates atavism analyzed by chi-squaretest
Characters / HKD / MR5 / Analisis P-Value / Conclusionobtained / table (α=5%)
Weight / 272,5 / 305 / 3,88 / 3,84 / significantly different
Horizontal circumference / 27 / 27,875 / 0,03 / 3,84 / insignificant
Vertical circumference / 26,125 / 28 / 0,13 / 3,84 / insignificant
Top diameter / 1,275 / 0,925 / 0,10 / 3,84 / insignificant
Bottom diameter (Turbin) / 1,675 / 1,325 / 0,07 / 3,84 / insignificant
Horizontal length / 8,15 / 8,75 / 0,04 / 3,84 / insignificant
Vertical length / 7,45 / 8,75 / 0,23 / 3,84 / insignificant
Outer skin thickness / 0,2375 / 0,175 / 0,02 / 3,84 / insignificant
Fruit meat thickness / 1,6 / 2,125 / 0,17 / 3,84 / insignificant
Total / 4,67 / 15,51 / insignificant
Results of the analysis in quantitative characters differences using the chi-square test at α = 5% (Table 3) shows that only the weight of melons which have significant different and there is no real difference to the other characters. This shows that the quantitative phenotypic characters from Hikadi and selection sample which experienced atavism has a high uniformity, or in other words quantitative morphological characters can be said to be uniform at α = 5% significance. It also can be said that the quantitative characters of Hikadi when compared with the results of sample that experienced atavism having fair uniformity, but qualitative phenotypic characters have significant differences. This could be due to the expression of genes that interact and bring back the characters of the parent derived from the interaction of paternally inherited genes and inherited through recessive pattern that cross-linked on the same chromosome; so when there were self-cross happened of F1 HK.10, these genes interacts and brings back the character Miyamauri-5 who have disappeared in 4 generations of Hikadi cultivars breeding. Thus, further molecular research that include similarity to other melon variety using RAPD (Random Amplified Polymorphic DNA) seems to be necessary to having full understanding of the observed atavism.
4. Conclusion
From the research that has been arranged, it can be concluded there were atavism occured from HK.10 self-breeding grown at Porok Beach, Kemadang, Gunungkidul. The morphological characters appears to be similiar with one of its ancestor from first generation, Miyamauri-5 (MR-5). The frequencies of plant that observed as atavism is 0,53% out of total population with 36.59% Ssm difference.
Acknowledgement
Authors would show the deepest gratitude towards Annas Rabbani for laboratory assistance and manuscript correction, Mr. Barno as local farmer at Gunungkidul, D.I. Yogyakarta, Indonesia for the support, and Indonesian Ministry of Finance for its support in this research through Riset Inovatif-Produktif Lembaga Pengelola Dana Pendidikan (RISPRO LPDP Kemenkeu) Research Grant No.: PRJ-457/LPDP/2014 and PRJ-622/LPDP/2016 for year 2014 to 2016.
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