
Cleomaceae Bercht. & J. Presl is a now a well-recognized family under Brassicales, placed in between Capparaceae and Brassicaceae (APG IV 2016).
The seeds of
The spatial distribution of genetic variation within plant populations is influenced by many factors such as gene flow, micro-environmental selection, spatial distribution pattern, life history, and human activities (Ying et al. 2007). The study of genetic diversity with molecular markers among the plant species has become crucial for understanding the genetic variation (Frascaroli et al. 2013). Screening of genetic variations among different species is essential in conservation genetics and molecular breeding techniques (Kalia et al. 2011; Shekhawat et al. 2018). Different molecular markers like Start Codan Targeted (SCoT), Inter-Simple Sequence Repeat (ISSR), Restriction Fragment Length Polymorphism (RFLP), Simple Sequence Repeats (SSR), Amplified Fragment Length Polymorphism (AFLP), and Random Amplified Polymorphic DNA (RAPD) are important tools for evaluating genetic variation among various genotypes (Frascaroli et al. 2013; Kudikala et al. 2020; Shekhawat et al. 2018). K’Opondo et al. (2009) assessed the genetic variation in the morphotypes of
The plant material of
The total DNA was isolated from the plants of the two varieties of
Three sets of markers were used for genetic variation studies of
List of SCoT, ISSR, and RAPD markers used in genetic study
S. No. | Primer Code | Primer Sequence (5–3) | Total number of alleles | Polymorphic alleles | Percentage polymorphism | PIC |
---|---|---|---|---|---|---|
1 | SCoT-1 | CAACAATGGCTACCACCC | 6 | 2 | 33 | 0.193 |
2 | SCoT-2 | CAACAATGGCTACCACCG | 1 | 1 | 100 | 0.032 |
3 | SCoT-3 | ACGACATGGCGACCAACG | 7 | 5 | 71 | 0.225 |
4 | SCoT-4 | CCATGGCTACCACCGCAG | 4 | 2 | 50 | 0.129 |
5 | ISSR-1 | GAGAGAGAGAGAGAGAG | 12 | 8 | 66 | 0.387 |
6 | ISSR-2 | GAGAGAGAGAGAGAGAT | 14 | 12 | 85 | 0.483 |
7 | ISSR-3 | AGAGAGAGAGAGAGAGTC | 14 | 10 | 71 | 0.451 |
8 | ISSR-4 | TGTGTGTGTGTGTGTGRA | 15 | 11 | 73 | 0.483 |
9 | OPA-1 | CAGGCCCTTC | 1 | 1 | 100 | 0.032 |
10 | OPA-2 | TGCCGAGCTG | 15 | 11 | 73 | 0.483 |
11 | OPA-3 | AGTCAGCCAC | 12 | 8 | 66 | 0.387 |
12 | OPA-4 | AATCGGGCTG | 1 | 1 | 100 | 0.032 |
Total | 102 | 72 | ||||
Average | 8.5 | 6 | 74 | 0.296 |
Each amplified product was scored either as 1 or 0, corresponding to the presence or absence of a band. The frequency of microsatellite polymorphism was calculated based on the presence or absence of common bands. The polymorphism information content (PIC) value = ∑ (1-Pi2)/n where ‘n’ is the number of band positions analyzed, and
The genus
The genus
The presence of two morphotypes of
Morphological distinction between the two varieties of
Character | var. |
var. |
---|---|---|
Habit | Annual erect herbs to <1 m tall | Annual erect herbs to 1.5 m tall |
Habitat | Terrestrial; fallow fields, dried-up ponds | Aquatic to semi-aquatic |
Leaflets: | ||
(i) Basal (proximal) | 7~9-foliolate, obovate, 3.5~12.2 cm long, entire | Same as in typical form, but bigger; 4-7 0.3~0.7 cm; crenate to crenate-serrate |
(ii) Middle (mesial) | 5~7-foliolate, elliptic to oblanceolate | 3~5 foliolate, elliptic lanceolate |
(iii) Upper (distal) | 1~5-foliolate, oblanceolate to lanceolate | 1~3; foliolate; often linear |
Internodes Pedicel length | Up to 5 cm 4 cm | Up to 15 cm 4.5 cm |
Flowers | 1.5~2.0 cm across | 2.5~3.5 cm across |
Petals | 4, close-by, elliptic, ovate to obovate; 1~1.4 × 0.3~0.8 cm; pink | 4, somewhat distant, oblong to ovate, 1.5~1.8 cm; pink |
Stamens (no/filaments) | ca. 35~60; 0.8~1.2 cm long | ca. 60~100 or more; 1.6~1.9 cm |
Capsule (beak length) | 2~4 mm | 5~10 mm |
Seeds | Ovoidal, 1.3~1.8 mm across; cleft more closed; testa tuberculate; appendages shorter, lesser, more or less pointed; with no waxy coating. | Rounded, 1.5~2.5 mm across; cleft more open; testa tuberculate with blunt tubercles of different heights; appendages with waxy coating. |
Three sets of markers (SCoT, ISSR, and RAPD) were used for genetic variation studies. The PCR analysis was performed with four primers each. Taxon–A (var.
The percentage of polymorphism was calculated according to the presence (1) or absence (0) of bands for SCoT, ISSR, and RAPD markers. The twelve primers used for genetic variation and diversity study generated 102 bands, an average 4.25 bands of SCoT, 14 bands of ISSR, and 7 bands of RAPD.
The genetic variation and differences were clearly seen between the two subpopulations of
The observed number of alleles (Na), effective number of alleles (Ne), gene diversity (H), Shannon’s information index (I), total heterozygosity (Ht), within-population heterozygosity (Hs), number of polymorphic bands (NPB) and Percentage of polymorphic bands (PPB) were calculated for the estimation of genetic variation between the two (A and B) subpopulations (Table 3).
Genetic diversity and differentiation parameters for Taxon A and B subpopulations of
Marker | Population | Diversity indices | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
na (± SD) | ne (± SD) | Np (Pl %) | h (± SD) | I (± SD) | Gd | Gi | Ht | Hs | Gst | Nm | ||
SCOT | Taxon–A | 1.225±0.425 | 1.161 ± 0.315 | 07 (22.58) | 0.092 ± 0.176 | 0.135 ± 0.256 | 0.0135 | 0.986 | 0.102 ± 0.026 | 0.096 ± 0.025 | 0.058 | 8.000 |
Taxon–B | 1.258±0.444 | 1.167± 0.297 | 08 (25.81) | 0.100 ± 0.175 | 0.149 ± 0.258 | |||||||
ISSR | Taxon–A | 1.548±0.505 | 1.432 ± 0.426 | 17 (54.84) | 0.237 ± 0.224 | 0.342 ± 0.319 | 0.0435 | 0.957 | 0.258 ± 0.049 | 0.241 ± 0.044 | 0.062 | 7.500 |
Taxon–B | 1.580 0.501 | 1.438± 0.408 | 18 (58.06) | 0.246 ± 0.217 | 0.356 ± 0.311 | |||||||
RAPD | Taxon–A | 1.354±0.486 | 1.238 ± 0.340 | 11 (35.48) | 0.141 ± 0.195 | 0.208 ± 0.286 | 0.0611 | 0.940 | 0.176 ± 0.026 | 0.151±0.020 | 0.142 | 3.000 |
Taxon–B | 1.387±0.495 | 1.283 ± 0.382 | 12 (38.71) | 0.161 ± 0.209 | 0.234 ± 0.302 | |||||||
ALL | ||||||||||||
(primers) | Taxon–A | 1.612± 0.495 | 1.323±0.341 | 19 (61.29) | 0.198 ± 0.187 | 0.303 ± 0.271 | 0.0133 | 0.986 | 0.208 ± 0.028 | 0.202 ± 0.026 | 0.025 | 19.255 |
Taxon–B | 1.741± 0.444 | 1.314± 0.278 | 23 (74.19) | 0.207 ± 0.159 | 0.328 ± 0.231 |
# value (mean) Taxon A:
Gd = Genetic distance; Gi = Genetic identity; Gst = Degree of genetic differentiation; h = Nei's (1973) gene diversity; Hs = Average heterozygosity; Ht = Heterozygosity at the polymorphic loci; I = Shannon's Information index (Lewontin 1972); na = Observed number of alleles; ne = Expected number of alleles/Effective number of alleles (Kimura and Crow 1964); Nm = Estimate of gene flow from Gst or Gcs. e.g., Nm = 0.5(1 - Gst)/Gst; Np = Number of polymorphic; Pl = Percentage of polymorphic loci; see McDermott and McDonald 1993).
(i) The Primers SCoT exhibited the genetic variation of Na (1.225 ± 0.425) Ne (1.161 ± 0.315), Np (07 (22.58)) h (0.092 ± 0.176) I (0.135 ± 0.256) for Taxon–A and Na (1.258 ± 0.444), Ne (1.167 ± 0.297), Np (08 (25.81)), h (0.100 ± 0.175), I (0.149 ± 0.258) for Taxon–B. (ii) The ISSR primers revealed the genetic variation as of Na (1.548 ± 0.50), Ne (5 1.432 ± 0.426), Np (17 (54.84)) h (0.237 ± 0.224) I ( 0.342 ± 0.319) in Taxon–A and Na (1.580 ± 0.501), Ne (1.438 ± 0.408), Np (18 (58.06)), h ( 0.246 ± 0.217), I (0.356 ± 0.311) for Taxon–B, and (iii) The RAPD primes have shown the genetic variation Na (1.354 ± 0.486), Ne (1.238 ± 0.340), Np (11 (35.48)), h (0.141 ± 0.195) I (0.208 ± 0.286) for Taxon–A in contrast to Taxon–B of Na (1.387 ± 0.495), Ne (1.283 ± 0.382) Np (12 (38.71)) h (0.161 ± 0.209) I (0.234 ± 0.302) (Table 3).
All the primers produced individual bands together for clear observation of genetic distance and variation between the two taxa as Na (1.612 ± 0.495), Ne (1.323 ± 0.341), Np (19 (61.29)), h (0.198 ± 0.187), I (0.303 ± 0.271) of Taxon–A and Na (1.741 ± 0.444), Ne (1.314 ± 0.27
Assessment of gene flow from one population to another is an essential parameter for the study of genetic diversity. Gene flow is a collective term that includes all the mechanisms resulting in the movement of genes from one species to another. In this study, lower relative differences between these two taxa 0.025 (Gst) indicates a high variation at a genetic level for these plants while the gene flow was showing high Nm value between them.
These results, as per SCoT, ISSR, and RAPD analyses, suggest that there is apparent variation within the species
The molecular data gathered in the present study of
The presence of the aquatic form of
1a.Plants terrestrial, of fallow fields; annual; erect herbs to 0.5 m height; leaflets elliptic to lanceolate; stamens about 60; distal leaves usually trifoliolate; seeds light brown, ovoidal, smaller (1.3~1.8 mm across); testa with tubercles pointed to curved, sparse; cleft somewhat closed·····································var.
1b.Plants aquatic to semi-aquatic, of littoral zones of water bodies; largely annual; erect to clump-forming herbs to 1.5 m height; stamens 60~100 or more; distal leaves linear; seeds dark brown, globose, bigger (1.5~2.5 mm across); testa with tubercles straight and blunt, denser; cleft fairly open·····································var.
Of the three kinds of molecular markers used to screen the intraspecific genetic variation in
SS is grateful to the University Grants Commission, New Delhi, for the award of BSR-RFSMS Fellowship. The authors are thankful to the Heads of the Departments of Botany and Biotechnology, Kakatiya University, Warangal, for facilities. VSR is obliged to Dr. Kanchi N. Gandhi, Senior Nomenclatural Registrar, Harvard University, Herbaria and Libraries, USA, for the useful discussion on nomenclature and to Dr. C. Murugan, BSI_SRC, for permission to work in MH.
The authors declare that they have no conflict of interest.
![]() |
![]() |