International Journal of Environment, Agriculture and Biotechnology

Vol-8, Issue-1; Jan-Feb, 2023

Journal Home Page Available: https://ijeab.com/

Journal DOI: 10.22161/ijeab

Peer Reviewed

Application of advanced molecular to select the variety of

Bitter gourd (Momordica charantia .L ) in Can Tho
Lang Thi Nguyen*, Hieu Chi Bui, Loan Hong Thi Nguyen, Phuoc Trong Nguyen

High Agricultural Technology Research Institute (HATRI); Web:hatri.org

*Corresponding author

Received: 27 Dec 2022; Received in revised form: 25 Jan 2023; Accepted: 02 Feb 2023; Available online: 09 Feb 2023
©2023 The Author(s). Published by Infogain Publication. This is an open access article under the CC BY license
(https://creativecommons.org/licenses/by/4.0/).

Abstract— A study was conducted to evaluate the genetic variation in bitter ground to using SNP (single-
nucleotide polymorphism) markers. Thirty -five primers showing reliable polymorphism were used .This
paper mainly applies the molecular directive from the self-absorbed population of F6 of Cho Moi/Ben Tre.
The Polymorphism on two SNP directives, TP1386 and TP 1877 with Bitter gourd on LG1. A wide variation
was observed for morphological traits like the number of days to the first male flower anthesis (29.33–33.67),
first female flower anthesis (30.5–38.6), fruit length (19.00–22.80 cm), fruit diameter (12.20–19.60 cm), and
yield per plant (933.8–1147.9 g).According to the GGT map, it is easier to determine the genetic pattern of
hybrids in the population compared to the genome of the parents in the F6 generation of the Cho Moi/Ben
Tre . With 34 SNPs (single-nucleotide polymorphism) molecules directives on LG1, the genetic distance
from 0-112. cM. The selected lines carried a superior homogeneity to the parent on the LG1. The result is 7
lines with F7 with 100% genes for hight yield the same with the father variety ( Ben Tre ), carrying hight
yield. The seven lines selected are: 1(F2-2-1-7-1); 2(F2-8-17-2-2); 5(F2 -5-3-1-5); 7(F2-25-15-8-7);
10(F2-54-4-1-10; 35(F2-10-6-5-35); 36(F2-5-2-7-36). However, after evaluating F7 lines and comparing
phenotypes and genotypes, there were only two lines: 2(F2-8-17-2-2); 7(F2-25-15-8-7)good appty for
breeding and hight yield . Named line 7(F2-25-15-8-7) was designated HATRI 07KQ . DNA Sequence of
HATRI 07KQ were submitted to GenBank .
Keyword— Bitter gourd , genetics and breeing , GGT, SNP (single-nucleotide polymorphism) molecular,
hight yield

I. INTRODUCTION is done mainly through hand pollination. Behera et al.

Bitter gourd (Momordica charantia L.; 2n = 22) is (2020) reported that F1 hybrid was derived from the crosses
an economically important vegetable crop belonging to the between pure-line of bitter gourd having good specific
subtribe Thalidianthinae, tribe Joliffieae, subfamily combiners for yield and its components. Valyaie et al.
Cucurbitoideae and family Cucurbitaceae (Jeffrey, (2021) reported that heterosis was obtained in seed quality
1980; De Wilde and Duyfjes, 2002). In bitter gourds, character and yield.
gynoecism is under the control of a single recessive gene But utilization of a gynoecious line would be more
(gy-1) (Ram et al., 2006; Behera et al., 2009; Matsumura et economical and easier method (Behera et al., 2009).
al., 2014), while two pairs of genes were reported by Cui et Conventional phenotypic selection for high and stable yield
al. (2018). The flowering traits like days to first pistillate requires the evaluation of yield in multiple environments
flower appearance, node at first pistillate flower appearance over several seasons; which is very expensive and time
and staminate: pistillate (♂:♀) flower ratio (sex ratio) are consuming (Yuan et al., 2002). Application of molecular
directly related to earliness and fruit yield. Production of indicator combining selection by traditional hybridization
hybrid seeds in bitter gourd is highly expensive because it method shortens time in breeding (Lang et al 2020).QTL

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Nguyen et al. Application of advanced molecular to select the variety of Bitter gourd (Momordica charantia .L )in Can Tho

analysis was performed for six major yield-contributing three replications, following recommended agronomic
traits such as fruit length, fruit diameter, fruit weight, fruit practices.
flesh thickness, number of fruits per plant and yield per DNA isolation and molecular marker analysis
plant. These six quantitative traits were mapped with 19
DNA extraction. The ninety varieties were grown in pots.
QTLs (9 QTLs with LOD > 3) using composite interval
Maximum protection was employed to ensure healthy and
mapping (CIM). Among 19 QTLs, 12 QTLs derived from
disease free-growth of seedlings. Leaves were collected 2-
‘Pusa Do Mausami’ revealed a negative additive effect
3weeks after planting for extraction of DNA.
when its allele increased trait score whereas 7 QTLs
derived from ‘DBGy-201’ revealed a positive additive Standard molecular grade chemicals and general
effect when its allele trait score increased( Rao et techniques for preparing stock solutions, buffers, reagents
al.,2021).The microsatellites i.e.SSR markers are mostly and equipment were followed according to Sambrook et al.,
preferred because of their co-dominance, repeatability and (1989). Molecular work was conducted at the Genetics and
easy transferability even though the initial cost of Plant Breeding Department of HATRI , Vietnam
development of these markers is very hig (Powell, et al DNA suitable for PCR analysis was prepared using
1996). However, the number of microsatellite markers a simplified procedure according to Mc Couch et al., (1988).
available in Momordica species is few(, Saxena, S. et al A piece of young rice leaf (2cm) was collected and placed
2015). It is established that a greater number of markers are in labeled 1.5ml centrifuge tube in ice. The leaf was ground
necessary for the development of a genetic map and marker- using a polished glass rod in a well of a Spot Test Plate
assisted selection( Tang et al.,2007). It is applied recently as (Thomas Scientific) after adding 400µl of extraction buffer
a very reliable tool for marker-assisted selection in .Grinding was done until the buffer turned green, an
accelerating crop improvement program ( He et al.,2014). indication of cell breakage and release of chloroplasts and
One major QTL qYD1 and two minor cell contents. Another 400µl of extraction buffer was added
QTLs qYD15 and qYD20 explained 23.28% of phenotyping into the well by pipetting. Around 400µl of the lysate was
variation for yield per plant. The QTLs identified in the transferred to the original tube of the leaf sample. The lysate
present study will be helpful in marker-assisted selection was deproteinized using 400µl of chloroform. The aqueous
and molecular breeding in bitter gourd crop improvement.( supernatant was transferred to a new 1.5ml tube and DNA
Rao et al.,2021) precipitated using absolute ethanol. DNA was air- dried and
In VietNam , very limited information is available for resuspended in 50µl of TE buffer ( Lang 2002)
determining nature of gene effects and inheritance of yield DNA quality checks used 1% agarose by melting
and yield contributing attributes in bitter gourd. For this 3g agarose in 300ml TAE buffer . The mixture was heated
purpose the present experiment was undertaken to breeding in microwave for 5-6 minutes and then cooled to around 55-
program with component and marker assited selection 600C. This was then poured on prepared electrophoresis box
(MAS )bitter gourd for the development of high yielding with combs. Gels were ready and combs removed after
variety. about 45 minutes. Seven microliliters of DNA sample plus
3µl loading buffer (Tris 1M pH = 8.0, glycerol, EDTA 0.5M
pH = 8.0, xylene cyanol 0.2%, bromphenol blue 0.2% and
II. MATERIALS AND METHODS
distilled water) was run at 70-80v, 60mA for 45 minutes or
Plant materials until loading buffer dye moved far from the wells. Gel was
A Cho Moi was crossed with Ben Tre of bitter gourd F7 then taken out and stained with ethidium bromide after
population was developed. Ten parents were crossed to which was visualized under UV light.( Lang 2002)
develop F1 seeds .Twenty of a single F1 plant derived from Microsatellite Analysis
the cross Cho moi/ Ben Tre were developed. The female
The whole microsatellite analysis
flowers of these plants were bagged before flowers opened
included PCR assay, polyacrylamide gel electrophoresis,
and were hand-pollinated by rubbing matured anthers of the
band detection and scoring.
male flowers on receptive stigma of the female flowers early
in the morning. The hybridized female flowers were kept PCR assay. Microsatellite primers were used to
bagged until formation of visually conspicuous green survey polymorphism on the samples. These were randomly
ovaries. The F1 plants selfed to develop F2 population (150), selected from the 34 microsatellite primer pairs currently
the F2 population was selfed individually to develop 150:F3 available for bitter groud(Rao et al., 2021). The PCR
families , plants selfed continue to F4: F5and F6 About 50 reaction was as follows:
F6:7 seeds from each family were sown in a single row with Reactions were overlaid with mineral oil and
processed in a Programmable Thermal Controller

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Nguyen et al. Application of advanced molecular to select the variety of Bitter gourd (Momordica charantia .L )in Can Tho

programmed for 35 cycles of 1 min at 94 0C, 1 min at 550C for aplot. Number of fruits per plant was calculated after
and 2 min at 720C, with a final extension at 750C for 5 min. dividing total number of fruits in a plot by total number of
After amplification, 10μl of stop solution was added to the plants in a plot.
PCR product which was then denatured at 940C for 2 min. GGT mapping assesses the genetic diversity of hybrid
Eight microliters of each reaction were run on populations
polyacrylamide gel.
(1) Genotype testing of hybrid populations on LG1
Band detection and scoring. Plates were separated using a is based on polymorphic molecular markers
plastic wedge and removed from the tank. The acrylamide between parent and parent plants. GGT
gel was soaked in ethidium bromide staining solution for 15 mapping assesses the genetic diversity of hybrid
to 20 minutes. Bands in the ethidium bromide-stained gels populations, thereby selecting individuals
were detected and photographed under UV light. Allelic carrying the desired target gene The GGT
bands were scored as 1 or 0 for presence or absence, method proposed by Tanksley et al. (1998) and,
respectively. Data were entered directly into an Excel Van Berllo (2008), Milne et al. (2010) built this
spreadsheet. useful software. GGT 2.0: "graphical
Data Analysis genotyping" is a new method developed by the
Analysis of variance. The agro-morphological data authors of Wageningen University, where
collected were initially analyzed through analysis of alleles express dominant contraction, recessive
variance to verify genetic variation in the traits measured. contraction, and heterozygosity in all hybrids in
The few traits with insignificant genetic variation, based on a population, allowing the selection of
the F-test, were not considered for further analyses. individuals to gather the desired genes in the
most effective way. GGT mapping method
Recording of trait data
through the following steps: Excel data file:
Fully matured unripe fruits were picked from each encoding the population gene with A, B being
F7 plant over the duration of fruit production. Some fruits the homozygous genotype of the parent tree; H
were studied in situ and left to ripen for collection of seeds. is a heterozygous genotype; Tumors are
Qualitative traits including fruit color were recorded unidentified genotypes; (2) Import data into the
visually. GGT window: convert Excel data to GGT data;
-Hight plant (m): The Hight of the plant was measured in (3) data processing in GGT.
meters from the ground level to the tip of the vine of plants III. 3. RESULT OF DISCUSSION

-Days to first male flowering: The number of days taken 3.1. Development of Bitter gourd hybrid
from sowing of seed to the opening of first male flower on populations
the plant was recorded as days to first male flowering and 3.1.1.Diversity of gene sources on parent Bitter
number of days taken from sowing of seed to the opening of gourd varieties from ( Cho Moi/ Ben Tre) The selection of
first female flower on the plant was recorded asdays to first peanut varieties using MAS has brought certain successes
female flowering. in recent times such as: shortening the selection time,
-Days to first female flowering: The number of days taken selecting varieties resistant to adverse conditions, disease-
from sowing of seed to the opening of first female flower on resistant varieties, quality varieties. Therefore, the parent
the plant was recorded as days to first female flowering. Bitter gourd varieties (Cho Moi and Ben Tre, ) were
genotype-analyzed to consider carrying hight yield genes,
-Fruit length (cm) and fruit diameter(cm): The observations
and at the same time, molecular markers of genes related to
regarding fruit length and diameter were measured from
yield and yield components. Sixty-nine molecular
five fruits, randomly selected from each treatment, at
directives were used to assess genetic diversity between
second, fourth and sixth pickings, respectively. Five
parent Bitter gourd varieties, but only 34 for
randomly selected fruits were taken from the harvested
polymorphisms with parents: including 2 molecular TP
fruits in each replication when
1386 and TP 1877 directives marked genes that regulate
it reached edible maturity. The fruits were cut longitudinally the hight yield .PCR production detected parent varieties
and length was measured with the help of a measuring tape ( with gene markers involved 16 marker TP 5296,
and fruit diameter was measured with digital Vernier TP5205,TP5058,TP3064,TP3003,TP2693,TP2480,TP2443
calipers. . TP2345. TP2313, TP2199,TP1992, TP1877, TP1459, TP
-Number of fruits per plant: Number of edible fruits was 1386 , TP 1323 on polyacrylamide gel with silver nitrate
counted at each picking and summed up for all the pickings staining( figure 1)

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Nguyen et al. Application of advanced molecular to select the variety of Bitter gourd (Momordica charantia .L )in Can Tho

M 12 12 12 12 12 12 12 12 12 1212 12 12 12 12 12

Fig.1: PCR production detected parent varieties ( with gene markers involved 16 marker TP 5296,
TP5205,TP5058,TP3064,TP3003,TP2693,TP2480,TP2443. TP2345. TP2313, TP2199,TP1992, TP1877, TP1459, TP 1386 ,
TP 1323 on polyacrylamide gel with silver nitrate staining.
Notes : M. DNA lamda , 1: Cho Moi and 2: Ben Tre.

3.1.2. Development of hybrid populations :The genotyped with markers, and the last four were identified as
parent seeds were planted at the HATRI Institute's green homozygous for both alleles with large particle sizes
house and resulted in 45 F1 plants. Twelve one plants (Figure 3, Figure 4, ). According to Rao et al. (2021), the
identified as hybrids actually carry both heterozygous nuclear indicator gene on Bitter gourd is controlled by the
alleles from their parents due to to molecular directives. hight yield localized on LG 1 associated with TP1877
These twelve one F1 are used to F2. Of the 150 F2 Plants, andTP 1459 , and TP1386. Three marker , TP1877 and TP
thirty-two were found to carry both parent alleles in 1459 , and TP1386 are therefore used to test genes
heterozygous condition. Generation F1, F2, F3, F6 were associated with hight yield .

Fig.2: Cho Moi and Ben Tre for parents and F7 generation hybrids: named HATRI 07KQ

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Nguyen et al. Application of advanced molecular to select the variety of Bitter gourd (Momordica charantia .L )in Can Tho

3.1.3. Application of molecular marker on F3 Marker TP1386 has a size (300-350bp)detected . In the F3
populations population with a common parent and the same population
Closely LG1 with marked by molecular marker size of 50 plants were also compared (Figure 3). The
TP1386. This gene is associated with the particle size group segregation pattern (Figures 3) . The results showed that
according to (Rao et al. (2021). The TP1386 molecular was there were 4 lines the same with Ben Tre ( 1,29,37 and 50 )
used on the F3 population to evaluate and select the high . and 20 lines the same with size marker Cho Moi (
yield of bitter ground. DNA is extracted from a segregating 14,15,16,24,25,26,27,28,30,31,32,33,34,35,36,37,38,40,41,
population derived from a cross of Cho Moi/ Ben Tre. 42( 300 bp) . Other individuals carry heterozygous
genotypes of the same size as their parents( 300-350bp).

Fig.3: PCR product amplified from DNA extract F3 population . The primers are TP 1386 with two bands position 300bp (
Cho Moi ) and 350bp ( Ben Tre ), on 3% agarose gel.
Note: M: molecure weight marker; P1: Cho Moi , P2: Ben Tre , 1-50 is F3

Similar to the results of the PCR product in 17 individuals in position 31,32

Figure 4 with the molecular directive TP 1877 recorded ,40,41,42,44,45,46,48,49,50 of the same size as Cho Moi
from the population Cho Moi/ Ben Tre give 6 homozygous with 250 bp. Other individuals carry heterozygous
plants with the allele of the product such as Ben Tre genotypes of the same size as their parents( 210-250bp).
variety.The results of the PCR product show that there are Thus, two molecular guidelines (TP 1877) on population
6 individuals at positions 1,2,5,7 ,10 and 26 of the same size show that the F3generation is still quite strongly dissociated
as Ben Tre corresponding to the size of 210bp. There are by 26-60% for the two molecular directives above in order.

Fig.4: PCR product of the molecular TP 1877 on 50 Lines of gene on LG 1, two bands position 250 bp ( Cho Moi ) and
210bp ( Ben Tre ), on 3% agarose gel.
Note: M: is the standard marker; P1: Cho Moi , P2: Ben Tre , 1-50 is F3

3.3. Selection of populations through GGT mapping The GGT map in Figure 5 shows that 7 individuals
3.3.1. Selection of F6 individuals of hybrid populations have 100% of the gene regions that coincide with the father
(Ben Tre), carrying the target gene hight yield . Selected
The F5 population of the Cho Moi/ Ben Tre hybrid
individuals are 1(F2-2-1-7-1); 2(F2-8-17-2-2); 5(F2 -5-3-1-
pair gives continued self-absorption and F6 generation
5); 7(F2-25-15-8-7); 10(F2-54-4-1-10); 35(F2-10-6-5-35);
selection. In the F6 population, 27 lines were selected for
36(F2-5-2-7-36). This noted that on 7 generation lines
genotype assessment through the chromosome LG1 map on
selected from F2 to F7, there was a homogeneity of large and
each GGT (Graphical genotyping), which is a method that
small particle sizes on these lines. Particularly, two
allows the expression of dominant, recessive, heterozygous
individuals 41(F2-7-2-9-41) recorded above accounted for
alleles of a population. The GGT map makes it easier to
95.2% of the large content of Ben Tre . This individuals
identify the genetic pattern of hybrids in the population
continue to choose the F8 generation to continues in
compared to the gene segment compared to the parent. GGT
breeding.
map is built on the Cho Moi/ Ben Tre platform in the F6
generation. On this map, the gene that regulates particle size 3.3.2.Phenotypic evaluation of the Cho Moi/ Ben Tre
traits is marked by 22 molecular indicators on LG1, the :the F6 population continued lines after evaluation by
migration distance. molecular directive continued planting for F7 generation
dissociation,particle size of selected lines in the field on the

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Nguyen et al. Application of advanced molecular to select the variety of Bitter gourd (Momordica charantia .L )in Can Tho

basis of the generation analyzed phenotype. In the (14.5-24.5 d) followed Days to first female flowering (19.5-
generations that continued to selected the F7generation, the 28.8 d). The length( 19-22.8 cm) and fruit diameter ( 12.2-
indicators hight plant, days to first male flowering, days to 19.6cm) .The yield of all the lines derived Cho Moi/Ben
first female flowering, fruit length, fruit diameter and Tre were hight yield to the yield of the standard checks with
number of fruits per plant were statistically significant. The parents . At Cai Răng , with eight F7 and two checks from
tallest variety was 2(F2-8-17-2-2) (231.5 cm) and the parents were evaluated in three replications . The highest
shortest was 10(F2-54-4-1-10) (165.3 cm). For Days to first yield was obtained from line 7(F2-25-15-8-7)
male flowering , the longest was of controlled parents trial (1147.9g/plant) next line 2(F2-8-17-2-2) ( 1145.8g/plant).

Fig.5: A. Generation F7 genetic diversity in hybrid populations of Cho Moi / Ben Tre on F7 of bitter grounder
Note: blue: genotype according to the parent ( Ben Tre ), red: genotype according to the parent plant ( Cho Moi ), gray:
heterozygous genotype, greencolor : marking selected individuals, 1-50: individuals of the Cho Moi/ Ben Tre population from
F6. Figure 5.B.
Table 1: Yield and component yield on bitter ground at CaiRang (CanTho)

Days to Days to Fruit length fruit

Hight first male first female (cm) diameter Number of
Bitter groud plant(cm) flowering flowering (Cm) fruits per plant
1(F2-2-1-7-1) 228,8b 21,2a 32,3c 21,1a 16,2c 1140,7b
2(F2-8-17-2-2) 231,5a 29,5a 38,67a 22,8a 15,8d 1145,8a
5(F2 -5-3-1-5) 209,9c 22,9a 32,9c 21,2a 19,8a 1140,5b
7(F2-25-15-8-7) 171,7d 22,2a 30,6c 22,2a 19,7a 1147,9a
10(F2-54-4-1-10) 165,3e 19,9b 35,6b 22,0a 16,5c 1138,4c

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Nguyen et al. Application of advanced molecular to select the variety of Bitter gourd (Momordica charantia .L )in Can Tho

35(F2-10-6-5-35) 227,8b 21,1b 35,9b 21,2a 13,5e 937,6d

36(F2-5-2-7-36) 222,2b 22,1b 37,8a 19,1b 12,4f 933,8d
41(F2-7-2-9-41) 221,2b 24,5c 30,5c 19,8b 12,2f 1128,0c
Cho Moi 226,5b 23,1b 38,8a 19,0b 15,4d 910,5d
Ben Tre 201,2c 16,1b 30,8c 20,9a 19,6a 1140,7b
Cv 9,2 1,1 4,9 1,22 1,38 2,62

IV. DISCUSSION regions that with the father ( Ben Tre variety ), carrying
The goal of a plant variety multiplication program hight yield . Selected individuals are two lines 7(F2-25-15-
is to make a change and select the desired genotype(s) for 8-7) (1147.9g/plant) next 2(F2-8-17-2-2) ( 1145.8g/plant)
cultivation or for breeding purposes. Selection of species apply in the future. Named line 7(F2-25-15-8-7) was
was based on careful consideration of a number of factors designated HATRI 07KQ . DNA Sequence of HATRI
including their economic importance in targeted 07KQ were submitted to GenBank .
geographical areas, nutrient density, access to genetic This noted that over 4 generation lines selected from F3to F7
resources, the improvement in the status of the crop there was a homogeneity of large and medium sizes on these
globally, and unsolved production constraints and lines. Molecular indicators are marked and developed
WorldVeg’s comparative advantage in solving them versus targeting marker loci associated with varieties for good
the private sector (World Vegetable Cente,2019). Bitter dissociation and high heterozygous rates are TP 1877 . The
ground , which are a highly self-pollinating crop, require hight yield and quality are important for bitter ground
special attention in detecting, passing high yields and selection and production, so a more mechanical
selecting because these processes require special skills and understanding of shell development and seed maturation
can be time consuming. Genetic research assists breeders in will be conducive to improving these characteristics.
understanding the mechanism of inheritance and improving
the effectiveness of a breeding program. Significant
V. CONCLUSION
progress has been made in the genetics and plant breeding
of peanuts for many years. In recent years, the discovery of Selected hybrid individuals must carry the
polymorphic molecular directives (SNPs) in combination dominant homologous gene on the corresponding
with developed sequencing technologies has led to a chromosome region obtaining 7 lines containing 100% of
significant improvement in fine mapping processes (Rao et the gene region that with the father (Ben Tre ), carrying the
al., 2021) .The yield of bitter ground is genetically target gene hight yield . A wide variation was observed for
controlled by polygenic factors. High-resolution mapping morphological traits like the number of days to the first male
of quantitative loci (QTLs) with linked markers can flower anthesis (29.33–33.67), first female flower anthesis
facilitate marker-assisted selection in seed selection for (30.5–38.6), fruit length (19.00–22.80 cm), fruit diameter
target characteristics. In the current study, with the (12.20–19.60 cm), and yield per plant (933.8–1147.9 g).
population of Cho Moi crossed with Ben Tre variety for With 34 SNPs (single-nucleotide polymorphism)
hight yield , there is an improvement. A graphical molecules directives on LG1, the genetic distance from 0-
representation of molecular marker data can be an important 112. cM. The selected lines carried a superior homogeneity
tool in the process of selection and evaluation of plant to the parent on the LG1. The result is 7 lines with 100%
material. A computer program was developed that enables genes for hight yield the same with the father variety ( Ben
representation of molecular marker data by simple Tre ), carrying hight yield. The seven lines selected are:
chromosome drawings in several ways. Commonly used 1(F2-2-1-7-1); 2(F2-8-17-2-2); 5(F2 -5-3-1-5); 7(F2-25-15-
marker file types that contain marker information serve as 8-7); 10(F2-54-4-1-10; 35(F2-10-6-5-35); 36(F2-5-2-7-36).
input for this program, which was named ‘GGT’ (an However, after evaluating F7 lines and comparing
acronym of Graphical GenoTypes)( phenotypes and genotypes, there were only two lines: 2(F2-
http://www.dpw.wau.nl/pv/pub/ggt/ www.plantbreeding.nl 8-17-2-2); 7(F2-25-15-8-7)good appty for breeding and
(in prep)2007. Besides representation, GGT can also be hight yield . Named line 7(F2-25-15-8-7) was designated
used for a diverse range of selections and analyses.Used to HATRI 07KQ . DNA Sequence of HATRI 07KQ were
build GGT maps based on the 34-loci molecular directive submitted to GenBank .
and extending the length of 112 cM. The GGT map in
Figure 5 shows that 7 individuals have 100% of the gene

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Nguyen et al. Application of advanced molecular to select the variety of Bitter gourd (Momordica charantia .L )in Can Tho

ACKNOWLEDGEMENTS [13] Yuan, J., Njiti, V. N., Meksem, K., Iqbal, M. J.,
Triwitayakorn, K., Kassem, M. A., et al. (2002). Quantitative
The authors would like to thank the Provincial People's
trait loci in two soybean recombinant inbred line populations
Committee and Can Tho Department of Science and segregating for yield and disease resistance. Crop Sci. 42,
Technology for providing funding to implement this 271–277. doi: 10.2135/cropsci2002.2710.
project. [14] VALYAIE, A., AZIZI, M., KASHI, A., SATHASIVAM, R.,
PARK, S.U., SUGIYAMA, A., MOTOBAYASHI, T. and
FUJII, Y., 2021. Evaluation of growth, yield, and biochemical
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