INDIAN JOURNAL OF PURE & APPLIED BIOSCIENCES

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Indian Journal of Pure & Applied Biosciences (IJPAB)
Year : 2020, Volume : 8, Issue : 3
First page : (37) Last page : (47)
Article doi: : http://dx.doi.org/10.18782/2582-2845.8096

RAPD Analysis of Genetic Diversity and Relationships among Kenaf (Hibiscus cannabinus L.) Germplasm

Mohammad Golam Mostofa* , Lutfur Rahman, Abu Saleh Muhammad Yahiya, Mohammad Harun-or-Rashid and Mohammad Mukul Mia
Breeding Division, Bangladesh Jute Research Institute, Manik Mia Avenue, Dhaka-1207, Bangladesh
*Corresponding Author E-mail: mostofabjri@gmail.com
Received: 15.04.2020  |  Revised: 21.05.2020   |  Accepted: 27.05.2020 

 ABSTRACT

To analyse the genetic diversity and relationships among kenaf germplasm, two cultivars and twenty three accessions from different geographical regions were assessed using RAPD markers. Six RAPD primers generated 38 polymorphic bands with an average of 6.33 polymorphic bands per primer. A high level of genetic variation (proportion of polymorphic loci 79.17%) and high genetic distance (0.0426 to 0.6523) were found among 25 indigenous and exotic kenaf germplasm. The UPGMA dendrogram based on Nei’s genetic distance segregated the genotypes into two major, two minor and one single genotype clusters. The distribution pattern of genotypes from different geographical regions into five clusters was random, indicating that geographical isolation may not be only factors causing genetic diversity. The minor cluster III comprised two accessions collected from same origin but they are distantly related to each other. The genetic distance between two cultivars was lower (0.0870) and they were found in the same cluster. These results indicate that studied kenaf germplasm not only exhibit a high level of genetic diversity but also have a different genetic background.  

Keywords: Diversity, Germplasm, RAPD, Kenaf

Full Text : PDF; Journal doi : http://dx.doi.org/10.18782

Cite this article: Mostofa, M.G., Rahman, L., Muhammad Yahiya, A.S., Harun-or-Rashid, M., & Mukul Mia, M. (2020). RAPD Analysis of Genetic Diversity and Relationships among Kenaf (Hibiscus cannabinus L.) Germplasm, Ind. J. Pure App. Biosci. 8(3), 37-47. doi: http://dx.doi.org/10.18782/2582-2845.8096

INTRODUCTION

Kenaf (Hibiscus cannabinus L.) is a fast growing annual plant that is harvested for its bast fibre. It is closely related to cotton and bhendi (H. esculentus) and is mostly grown over a wide range of latitude from 16oS to 41oN (Kumar, 1999). Kenaf, a jute substitute, has capacity to produce a huge amount of biomass and hence it is considered presently as the main renewable source of raw materials for paper pulp production in many countries of the world (Wood, 1981; Hazra & Singh, 1997; Sinha and Day, 2008). Being tolerant to moisture stress, it can be grown in drought-prone areas and in low fertile soils where jute cannot be grown. It is a plant all parts of which have extensive uses.

Kenaf fibres are used by mills for making cordage, yarn, sacks and hessian cloth and also in combination with other synthetic fibres or natural fibres like jute (Maiti, 1997). Its leaves are a part of human diet in some parts of India and Africa (Killinger, 1969). Kenaf twigs and seeds are good feed for milch cows and dry stem is used for fuel, fencing, match sticks, climbing sticks of vegetables field, cattle shed and other domestic purposes. Moreover, it is biodegradable and environment friendly crop.
               Kenaf is believed to be originated in Africa, more particularly in East Africa, where it is found in wild form. It was domesticated in Indian sub-continent as bast fibre crop. Later on, it is distributed throughout the tropical and sub-tropical parts of the world (Karmakar et al., 2008). Kenaf has a good potential of becoming an excellent source of various products such as poultry litter and cattle feed (Perry et al., 1993). It can be processed into acoustic tile, animal bedding, soil-less potting mixes, composite board for construction, mats for erosion control and absorbents for cleaning up chemicals or oils spills (Banuelos, 2000). Kenaf sticks contain around 38% cellulose, 25% hemi-cellulose and 20% lignin (Pandey & Krishnan, 1990). Being cheap and easily available in large quantities, it is an ideal ligno-cellulosic substrate for producing industrial raw materials for much higher values. Kenaf seeds contain about 21% oil, which has various industrial uses like manufacture of soaps, linoleum paints and for lubricating purposes.
            Kenaf can absorb CO2 and NO2 3-5 times faster than forests. It can clean the environment efficiently (Lam, 2000). In some Japanese cities, kenaf was planted by government to improve the air quality. In Bangladesh, kenaf is one of the most important bast fibre crops and the area under its cultivation is increasing day by day. Around 50 thousand hectares of land is now being under kenaf cultivation and the fibre production is 100-110 thousand tons per annum with average yields of 2.0-2.5 tons/ha. From the beginning of crop improvement program in Bangladesh on kenaf, four high yielding varieties viz. HC-2 (popularly known as Joli kenaf), HC-95, BJRI Kenaf-3 and BJRI Kenaf-4 were developed through pure line selection from a few common accessions. These varieties, however, have some undesirable characters. They contain very limited genetic variability with respect to adaptability to different agro-climatic zones, fibre yield and quality. They are susceptible to root-knot nematode (Meloidogyne spp.), spiral borer (Agrilus acutus), yellow mosaic disease and have prickles on stems and bristles on fruits (a character disliked by farmers due to irritating to the skin). These undesirable characters limit their large-scale adaptation and extension. Therefore, systematic collection, characterization and utilization of diverse kenaf genetic resources are needed to overcome such problems.
            Genetic diversity is essential to meet the diverse goals such as producing cultivars with increased yield (Joshi & Dhawan, 1986), wider adaptation, desirable quality, pest and disease resistance (Nevo et al., 1982). For the study of genetic diversity, the plant scientists have used traditionally morphological and physiological features of plants. But in most cases, plant genomes have large amounts of repetitive DNA which are not expressed and do not contribute to the physiological or morphological appearance of the plants. In the case of very closely related plant varieties and species, there are very few morphological differences, which as a matter of fact, do not represent the true genetic differences at the DNA level. So, there is always a need to study polymorphism at DNA level, which can be indicative of genetic diversity. RAPD technology has been used for assessing germplasm for species identification (Welsh & McClelland, 1990) and molecular characterization of many crop species, such as soybean, rice, rose and mustard (Fujishiro & Sasakuma, 1994). Similarly, varietal identification of kenaf has been done through RAPD (Rahman etal., 2007) but large scale germplasm characterization has yet been rare. Zhou et al. (2002) studied with 14 kenaf varieties and reported that identification of individual kenaf variety through morphological characterization was difficult but it was clearly separated by RAPD analysis. They concluded that RAPD analysis is an effective tool for identifying kenaf varieties and determining their genetic relationship to a certain extent.
            In this study, we investigated the genetic diversity and relationships among kenaf accessions and cultivars based on RAPD markers, and also examined its efficiency in the light of the known origin of the study materials.

MATERIALS AND METHODS

Plant materials
Twenty three kenaf accessions and two cultivars were obtained from the Genebank of Bangladesh Jute Research Institute (BJRI), Dhaka (Table 1). These germplasm were collected from different geographical regions (16 countries) and coded herein G1 through G25. The two cultivars – HC-2 and HC-95 are widely used as commercial variety and also used as parental material for kenaf breeding program in Bangladesh.

Table 1: Kenaf accessions and cultivars analysed

Sl. No.

Country of origin

Accession and cultivar name

01

Bangladesh

G1 (HC-2), G2 (HC-95), G4 (Acc. 2731), G10 (Acc. 2103)

02

Java

G3 (Acc. 1993)

03

Netherlands

G5 (Acc. 2922)

04

Thailand

G6 (Acc. 4634)

05

Australia

G7 (CPI-72126), G8 (Acc. 4659)

06

USA

G9 (Acc. 4718), G14 (Acc. 4628), G23 (Acc. 4750)

07

Poland

G11 (Acc. 4372)

08

Sudan

G12 (Acc. 4383), G21 (Acc. 4389)

09

El-Salvador

G13 (Acc. 4410)

10

China

G15 (Acc. 4922), G24 (Acc. 5017)

11

Pakistan

G16 (Acc. 5050), G25 (Acc. 5030)

12

Nepal

G17 (Acc. 5080)

13

Cuba

G18 (Acc. 1983)

14

Kenya

G19 (Acc. 4197)

15

Tanzania

G20 (Acc. 4348)

16

France

G22 (Acc. 4432)

DNA extraction and RAPD analysis
Total genomic DNA was isolated from a bulk of young leaf tissues from five plants grown in greenhouse, following the modified Doyle and Doyle’s (1990) protocol of CTAB (Cetyltrimethyl ammonium bromide) method (Murray and Thompson, 1980). The experiment was carried out at Genetic Fingerprinting Laboratory of the Department of Genetics and Plant Breeding, and Central Laboratory of Bangladesh Agricultural University, Mymensingh. Nineteen primers of random sequence were screened on a sub sample of 4 randomly chosen individuals from 25 different kenaf genotypes to evaluate their suitability for amplifying DNA sequences that could be accurately scored. Primers were evaluated on the basis of band resolution or intensity, repeatability of markers, presence of smearing, consistency within individuals and the potential to differentiate polymorphism. The details of the primers are given in Table 2. A final subset of 6 primers exhibiting good quality banding patterns and sufficient variability were selected for the analysis of whole sample set of genotypes.

Table 2: List of random primers used in the study for screening

Primer code

Sequence (5' - 3')

GC content (%)

GLA05

AGGGGTCTTG

60

GLA07

GAAACGGGTG

60

GLA09

GGGTAACGCC

70

GLA10

GTGATCGCAG

60

OPG1

TGCCGAGCTG

70

OPG2

AGTCAGCCAC

60

OPG3

GAAACGGGTG

60

OPG4

GGGTAACGCC

70

OPG5

GTGATCGCAG

60

OPG6

CAATCGCCGT

60

OPG7

CAGCACCCAC

70

OPG8

CCGCCCAAAC

70

OPG9

AGCGAGCAAG

60

OPG10

GAACACTGGG

60

OPG11

CCCTACCGAC

70

OPG12

AATGGCCCAG

60

OPG13

CTCCTGCCAA

60

OPG14

CCCAGCTGTG

70

OPG15

GTGTCGCGAG

70

Primers marked with bold letter and underline showed polymorphism and were selected for RAPD analysis

DNA amplification was conducted as described by Williams et al., (1990) with some modifications. PCR reactions were performed in a 25µl reaction mix consisting of
■  Taq DNA polymerase buffer (10 x) = 4.00µl
■  Primer (10µM)                           = 3.00µl
■  dNTPs (250µM)                        = 0.50µl
■  Taq DNA polymerase               = 0.20µl (1 unit)
■  Genomic DNA (25 ng/µl)         = 3.00µl
■  Sterile deionized water              = 14.30µl
Amplification was carried out in a thermal cycler (Master Cycler Gradient, Eppendorf ) that was programmed as an initial denaturation or preheating at 94 oC for 3 minutes (first cycle), followed by 40 cycles of the procedure at 94 oC for 1 minute (denaturation), 36 oC for 1 minute (annealing), 72 oC for 2 minutes (extention/elongation), and adding a final elongation at 72 oC for 5 minutes (last cycle). After completion of cycling program, reactions were held at 4 oC followed by cooling. The PCR products were then separated electrophoretically on 1.5% agarose gel containing ethedium bromide in 1 X TBE buffer at 100V for 1 hour. DNA bands were observed under UV-light on a Transilluminator and photographed by a Gel Cam Polaroid camera (Type 667).
Data analysis
Polymorphic RAPD markers were manually scored as binary data: present (1) or absent (0) for each individual and each primer. Only clearly distinguishable bands were scored. The scores obtained using all primers in RAPD analysis were then pooled for constructing a single data matrix. This was used to estimate polymorphic loci, Nei’s (1973) gene diversity and Nei’s (1972) genetic distance (D) using a computer program, POPGEGE (version 1.31) (Yeh et al., 1999). To investigate the genetic relationships among accessions, genetic distances between all pairs of individual accession were estimated. A dendrogram was constructed based on genetic distance using unweighted pair group method with arithmetic averages (UPGMA).  

RESULTS AND DISCUSSION

Nineteen arbitrary decamer primers were initially surveyed for their ability to produce polymorphic patterns. Out of them, six primers viz. GLA09, GLA10, OPG1, OPG2, OPG7 and OPG9 yielded comparatively higher number of amplification products with higher intensity, minimal smearing and good resolutions with clear bands. Hence, these six primers were selected for evaluation of diversity across all the genotypes. Amplification of the isolated genomic DNA from each of 25 germplasm using selected primers revealed a variety of RAPD patterns (Plate 1). Clearly detectable and reproducible bands ranged from 220 to 850 bp and 270 to 650 bp in size in case of primer GLA09 and OPG1. A total of 48 scorable bands produced from six primers of random sequence and their size ranges (bp) are presented in Table 3.

Table 3: RAPD primers with corresponding bands score and their size range together with polymorphic bands observed in 25 kenaf genotypes

Primer code

Sequences
(5´-3´)

Total number of bands scored

Size ranges (bp)

Number of polymorphic bands

GLA09

GGGTAACGCC

10

220 – 850

8

GLA10

GTGATCGCAG

9

210 – 1200

7

OPG1

TGCCGAGCTG

7

270 – 650

7

OPG2

AGTCAGCCAC

7

180 – 1800

5

OPG7

CAGCACCCAC

6

270 – 1500

3

OPG9

AGCGAGCAAG

9

330 – 800

8

   Total

48

 

38

   Average

8

 

6.33

Lane

Genotypes

Lane

Genotypes

Lane

Genotypes

Lane

Genotypes

Lane

Genotypes

1

HC-2

6

Acc. 4634

11

Acc. 4372

16

Acc. 5050

21

Acc. 4389

2

HC-95

7

CPI-72126

12

Acc. 4383

17

Acc. 5080

22

Acc. 4432

3

Acc. 1993

8

Acc. 4659

13

Acc. 4410

18

Acc. 1983

23

Acc. 4750

4

Acc. 2731

9

Acc. 4718

14

Acc. 4628

19

Acc. 4197

24

Acc. 5017

5

Acc. 2922

10

Acc. 2103

15

Acc. 4922

20

Acc. 4348

25

Acc. 5030

The efficiency of molecular marker technique depends upon it polymorphism level in the set of accessions tested. In this study, RAPD markers amplified a total of 48 bands of which 38 (79.17%) were found to be polymorphic and the rest 10 (20.83%) were monomorphic in nature. On an average, each primer generated 6.33 polymorphic bands (Table 3). This is in accordance with the findings of Geo-Anping et al. (2002) who worked with different Hibiscus species using RAPD primers and showed that 16 primers of arbitrary sequence out of 80 amplified 192 bands of which 149 bands were polymorphic and 43 were monomorphic.
The number of amplified fragments against genotypes and primers varied widely. Size of PCR amplification products scored for each individual of 25 kenaf genotypes, and for each primer is presented in Table 4.

Table 4: Number of amplified fragments scored against genotypes and primers

 Genotypes

Primers

GLA 09

GLA 10

OPG 1

OPG 2

OPG 7

OPG 9

Total band

 

Polymorphic band

  G1 (HC-2)

3

5

2

4

4

3

21

11

  G2 (HC-95)

4

5

3

6

4

3

25

15

G3 (Acc. 1993)

6

5

2

3

4

3

23

13

G4 (Acc. 2731)

7

4

2

3

5

4

25

15

G5 (Acc. 2922)

7

5

2

5

4

4

27

17

G6 (Acc. 4634)

7

5

2

4

5

4

27

17

G7 (CPI-72126)

7

6

3

6

3

2

27

17

G8 (Acc. 4659)

7

5

4

5

4

3

28

18

G9 (Acc. 4718)

9

7

2

5

5

5

33

23

G10 (Acc. 2103)

6

7

3

5

5

5

31

21

G11 (Acc. 4372)

8

4

5

5

4

3

29

19

G12 (Acc. 4383)

8

6

2

5

4

2

27

17

G13 (Acc. 4410)

8

6

4

5

4

4

31

21

G14 (Acc. 4628)

2

6

4

6

3

5

26

16

G15 (Acc. 4922)

7

6

4

6

4

5

32

22

G16 (Acc. 5050)

8

4

2

5

3

3

25

15

G17 (Acc. 5080)

8

3

4

6

4

3

28

18

G18 (Acc. 1983)

6

6

4

5

4

4

29

19

G19 (Acc. 4197)

6

7

2

6

4

4

29

19

G20 (Acc. 4348)

7

6

2

6

4

5

30

20

G21 (Acc. 4389)

6

5

2

5

4

5

27

17

G22 (Acc. 4432)

6

4

4

6

4

4

28

18

G23 (Acc. 4750)

6

4

3

5

4

4

26

16

G24 (Acc. 5017)

8

4

4

5

5

2

28

18

G25 (Acc. 5030)

8

4

4

5

4

3

28

18

Total

165

129

75

127

102

92

690

440

The primer GLA09 is able to produce a total of 165 bands in 25 genotypes (average 6.6 bands per genotype) and the highest number (9 bands) was recorded in G9 (Acc. 4718). On the contrary, the lowest number of bands (75 or 3 bands/genotype) was recorded in primer OPG1. From the polymorphic loci point of view, G9 produced the highest number of polymorphic bands (23) across all primers and G1 produced the least (11 polymorphic bands). It means, none of the primers alone was able to identify all genotypes. Similar results were obtained by Vilarinhos et al. (2000) in Citrus, who identified 12 hybrids with six of 20 primers tested. None of 20 primers used by them was useful singly to identify all 12 hybrids.
The values of Nei’s (1973) gene diversity and Shannon’s information index for different kenaf genotypes across all loci are shown in Table 5. The estimate of Nei’s (1973) genetic diversity for entire genotypes was 0.2118 and Shannon’s information index was 0.3363.

Table 5: Estimation of genetic variability

No. of polymorphic loci

Proportion of polymorphic loci
( %)

Nei’s (1973) gene diversity
( h )

Shannon’s information index
( i )

38

79.17

( 0.2118 ± 0.1668 )

( 0.3363 ± 0.2349 )

The high level of polymorphism revealed by the proportion of polymorphic loci (79.17%) indicated that there was a high level of genetic variation among the studied genotypes. Estimates of Nei’s (1973) gene diversity (0.2118) and Shannon’s information index (0.3363) across all loci (Table 5) also support the existence of high level of genetic variation in all studied materials.
The amplified products were scored and used for construction of a dendrogram (Figure 1) as well as for determining their genetic distances. Table 6 shows the distance between kenaf genotypes. The values of pair-wise comparison of Nei’s (1972) genetic distance (D) between genotypes computed from combined data sets for six primers ranged from 0.0426 to 0.6523. 

Table 6: Five of each higher and lower Nei’s (1972) genetic distance (D) between pairs of genotype based on RAPD markers

Five higher
D values

Genotype combination

Five lower
D values

Genotype combination

0.6523

G14 x G10, G23 x G10

0.0426

G25 x G17, G25 x G24

0.5754

G11 x G10, G15 x G9, G17 xG10

0.0645

G21 x G20

0.5390

G10 x G1, G10 x G4

0.0870

G2 x G1, G3 x G1, G4 x G3, G5 x G4, G6 x G4, G6 x G5, G18 x G5, G19 x G5, G19 x G18, G24 x G11, G24 x G17, G25 x G11, G25 x G16

0.5039

 G10 x G8, G14 x G9, G16 x G10, G22 x G10, G24 x G10, G25 x G10

0.1100

G16 x G5, G16 x G12, G24 x G12, G25 x G5, G25 x G12, G25 x G18

0.4700

G10 x G3, G10 x G5, G10 x G9, G12 x G10, G20 x G14, G23 x G20

0.1335

G3 x G2, G5 x G3, G6 x G3, G7 x G5, G12 x G3, G12 x G5, G13 x G7, G16 x G11, G17 x G11, G17 x G16, G18 x G7, G18 x G13, G19 x G3, G19 x G6, G19 x G9, G22 x G8, G22 x G11, G22 x G17, G24 x G16, G24 x G22, G25 x G22

The highest genetic distance (0.6523) was observed between the genotypes G10 and G14, and G10 and G23. While the lowest genetic distance (0.0426) was observed between the genotypes G17 and G25, and G24 and G25. The distance between the highest and the lowest values indicated the presence of variability among 25 kenaf genotypes. The dendrogram shows five different clusters designated as I, II, III, IV and V, in which cluster I, II and IV were further divided into ten sub-clusters. The distribution of cluster members is shown in Table 7.

Table 7: Distribution of 25 kenaf genotypes under different clusters using RAPD data

Cluster number

Total no. of genotypes in

Genotypes included in different clusters

Cluster

Sub-cluster

I

8

Sc1 = 2

G1(HC-2), G2(HC-95)

ScII = 4

G3(Acc.1993), G4(Acc. 2731), G5(Acc. 2922), G6(Acc. 4634)

ScIII = 2

G18 (Acc. 1983), G19 (Acc. 4197)

II

10

ScI = 2

G7 (CPI-72126), G13 (Acc. 4410)

ScII = 2

G8 (Acc. 4659), G22 (Acc. 4432)

ScIII = 4

G11(Acc. 4372), G17(Acc.5080), G24(Acc. 5017), G25(Acc.5030)

ScIV = 2

G12 (Acc. 4383), G16 (Acc. 5050)

III

2

-

G14 (Acc. 4628), G23 (Acc. 4750)

IV

4

ScI = 1

G10 (Acc. 2103)

ScII = 1

G15 (Acc. 4922)

ScIII = 2

G20 (Acc. 4348), G21 (Acc. 4389)

V

1

-

G9 (Acc. 4718)

Dendrogram based on Nei’s (1972) genetic distance using UPGMA indicated segregation of 25 kenaf genotypes into two major, two minor and one single genotype clusters. Identical and closely related genotypes were clustered together. The last single one genotype cluster appeared as outlier in the dendrogram and distantly related with the rest of the genotypes. The minor clusters had 2 to 4 genotypes. The two major clusters consisted of 8 to 10 genotypes. The first major cluster formed three separate sub-clusters and the second major cluster was subdivided into four sub-clusters (Table 7). The first minor cluster composed of two genotypes and the second minor cluster grouped together four genotypes. Similarly, Suvakanta et al. (2006) studied with two species of Hibiscus (H. sabdariffa and H. schizopetalus) and 16 varieties of Hibiscus rosa-sinesis through RAPD markers and reported that the studied materials formed a single cluster. The first major cluster consisted of three varieties; and a second major cluster consisted of two species and 13 varieties. The genetic distance was very close within the varieties and also among the species.

CONCLUSION

Diversity analysis is usually performed to identify the diverse genotypes for hybridization purposes. The genotypes grouped together are less divergent among themselves than those which are fall into different clusters. The crosses involving parents belonging to the maximum divergent clusters were expected to manifest maximum heterosis in F1 and wide variation in F2. In choosing parental materials for crossing work, selection criteria should be based on genetic distances between genotypes and information on their relationship in cluster analysis. Cox et al. (1985) proposed that crosses between distantly related lines in an inbred improvement program would increase the number of segregating loci in the F2 and subsequent inbred generations. Cross combinations involving parents that are distantly related and coming from different clusters are more expected to produce heterotic offspring. 
Molecular identity of kenaf genotypes, particularly for the released varieties, is very important to protect bio-piracy. In the present study, twenty five genotypes of kenaf were used for RAPD analysis using six decamer random primers. Amplification of the isolated genomic DNA from each of the 25 genotypes, revealed a variety of RAPD patterns. A total of 48 RAPD markers were generated of which 38 (79.17%) were considered as polymorphic and the rest were monomorphic in nature. Primers GLA09 and OPG9 individually amplified the maximum number of polymorphic bands (8) and the minimum number (3) was recorded with primer OPG7. On an average each primer generated 6.33 polymorphic bands. The estimate of Nei’s (1973) genetic diversity for 25 kenaf genotypes was 0.2118 and Shannon’s information index was 0.3363 across all loci indicated the presence of high level of genetic variation among the studied genotypes.
The UPGMA dendrogram based on Nei’s (1972) genetic distances indicated segregation of 25 kenaf genotypes into two major, two minor and one single genotype clusters. The first and second major cluster consisted of 8 and 10 genotypes, respectively; while the two minor clusters (III and IV) had 2 to 4 genotypes. The highest genetic distance (0.6523) was found between G10 and G14 and/or between G10 and G23 and they remain in different cluster though G14 and G23 formed same cluster. Likewise, the genetic distance between G17 and G25 and/or between G24 and G25 was lowest (0.0426) and they remain together in the same cluster.
However, the relationships observed using the RAPD-based dendrogram may provide detail information on the studied materials. Considering interrelationships and genetic distances the genotypes G4, G5, G7, G8, G19 and G25 may be selected as parental source for future breeding program of kenaf improvement.

Acknowledgements

The authors thank the Bangladesh Jute Research Institute (BJRI) for supplying the valuable seeds of kenaf accessions and cultivars used in this research. This research work was fully supported by the Bangladesh Agricultural Research Council, Ministry of Agriculture, Bangladesh. The authors expressed their deepest gratitude and appreciation to Dr. Asma Khatun, Dr. M. M. Hussain, Dr. C. K. Saha, Dr. M. S. Haque, Dr. M. S. Islam; and Director Agriculture of BJRI Dr. M. M. Rahman for their inspiration and kind support. The authors also expressed their heartiest gratitude to all personnel of Breeding Discipline, BJRI for their contribution regarding this study.

REFERENCES

Banuelos, G.S. (2000). Kenaf and Canola–Selenium Slurpers. Agril. Res., June 2000: 10-11.
Cox, T.S., Lookhart, G.L., Walker, D.E., Harrell, L.G., Albers, L.D., & Rodgers, D.M. (1985). Genetic relationships among hard red winter wheat cultivars as evaluated by pedigree analysis and gliadin polyacrylamide gel electrophoretic patterns. Crop Sci., 25, 1058-1063.
Doyle, J. J., & Doyle, J.L. (1990). Isolation of plant DNA from fresh tissues. Focus, 12, 13-15.
Fujishiro, T., & Sasakuma, T. (1994). Variety identification and molecular characterization of newly bred line by RAPD marker in Brassica juncea. Breed. Sci., 44(1), 132.
Geo-Anping, Zhou-Peng, Su-Jian Gusng, Guo-AP, Zhou-P and Su-JG (2002). Random amplified polymorphic DNA (RAPD) analysis among Hibiscus cannabinus and related species. J. Tropic., & Sub-tropic. Bot., 10(4), 306-312.
Hazra, S.K., & Singh, D.P. (1997). Paper making from bast fibre crops- Aspects and Prospects. Technical Bulletin Series No.4. CRIJAF, p.56.
Joshi, A.B., & Dhawan, N.L. (1986). Genetic improvement of yield with special reference to self fertilizing crops. Indian J. Genet., 26(1), 101-113.
Karmaker, P.G., Hazra, S.K., Ramasubramanian, T., Mandal, R.K., Sinha, M.K., & Sen, H.S. (2008). JUTE AND ALLIED FIBRE UPDATES: Production and Technology. Central Research Institute for Jute and Allied Fibres (ICAR), Barrackpore, Kolkata-700 120, India. www.crijaf.org
Killinger, G.B. (1969). Kenaf (Hibiscus cannabinus), a multi-use crop. Agron. J., 61(5): 734-736.
Kumar, D. (1999). Possibility of commercial utilization of residual heterosis in kenaf (Hibiscus cannabinus L.). Indian J. Genet., 59(2),227-232.
Lam, T.B.T. (2000). Structural details of kenaf cell walls and fixation of carbon dioxide. Proceedings of the 2000 international kenaf symposium. Hiroshima, Japan, Oct.13-14, pp. 81-90.
Maiti, R.K. (1997). World Fiber Crops. Science Publishers Inc. New Hampshire, USA. pp. 41-61.
Murray, M.G., & Thompson, W.F. (1980). Rapid isolation of high molecular weight plant DNA. Nucl. Acids Res., 8, 4321-4325.
Nei, M. (1972). Genetic distance between populations. Am. Nat., 106, 283-292.
Nei, M. (1973). Analysis of gene diversity in subdivided populations. Proc. Natl. Acad. Sci., 70, 3321-3323.
Nevo, E., Golenberg, E., Beilies, A., Brown, A.H.D., & Zohary, D. (1982). Genetic diversity and environmental association of wild wheat, Triticum diococcoides in Israel. Theor. Appl. Genet., 62, 241-254.
Pandey, S.N., & Krishnan, A. (1990). “Fifty years of research (1939-1989)”, Jute Technological Research Institute, p.80.
Perry, R.C., Jones, D.E., & Bhangoo, M.S. (1993). “A preliminary study on kenaf as a feed for livestock (1992)”, pp. 45-48 in proceedings. Fifth Annual International Kenaf Association Conference, March 3-5, 1993, Fresno CA.
Rahman, L., Molla, M.R., Sultana, S., Islam, M.N., Ahmed, N.U., Rahman M.S., & Nazim-ud-Dowla, M.A.N. (2007). Plant Varieties of Bangladesh: Morphological and molecular characterization. Published by Seed wing, MoA, GoB, Khamarbari, Dhaka. 1, 184-215.
Sinha, M.K., & Day, A. (2008). Pulp and Paper from Jute and Allied Fibres. In: P.G. Karmakar, S.K. Hazra, T. Ramasubramanian, R.K. Mandal, M.K. Sinha and H.S. Sen (ed), JUTE AND ALLIED FIBRES UPDATES: Production and Technology. CRIJAF, India. pp. 287-296.
Suvakanta, B., Senapati, S.K., Subhashree, A., Anuradha, M., & Rout, G.R. (2006). Identification and genetic variation among Hibiscus species (Malvaceae) using RAPD markers. Biosciences, 61(1/2), 123-128.
Vilarinhos, A.D., Viana, C.H.P., Soares, F.W.S., Nickel, O., Oliverra, R.P., & de Oliveira, R.P. (2000). RAPD markers for evaluating genetic diversity and identifying inter-specific hybrids in citrus. Revista Brasileira de Fruticultura, 22(21), 14-19. [Cited from CAB Abst., Vol. CIAC, 2000-2002].
Welsh, J., & McClelland, M. (1990). Fingerprinting genomes using PCR with arbitrary primers. Nucl. Acids Res., 18: 7213-7218.
Williams, J.G.K., Kubelik, A.R., Livac, K.J., Rafalski, J.A., & Tingey, S.V. (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res., 18(22), 6531-6535.
Wood, I.M. (1981). The utilization of field crops and crop residues for paper pulp production. Fields Crops Abstract. Commonwealth Bureau of Pastures and Field Crops, 34(7): 557-568.
Yeh, F.C., Yang, R.C., Boyle, T.B.J., Ye, Z.H., & Mao, J.X. (1999). POPGENE, the user-friendly software for population genetic analysis. Mol. Biol. Biotechnol. Centre, University of Alberta, Canada.

Zhou Cheng, Bao-Rong Lu, Brian S. Baldwin, Kazuhiko Sameshima and Jia-Kuan Chen (2002). Comparative studies of genetic diversity in kenaf (Hibiscus cannabinus L.) varieties based on analysis of agronomic and RAPD data. Hereditas, 136, 321-239.

 

 

 

 

 

 

 




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