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\begin{document}

\title{STUDIES ON BAMBARA GROUNDNUT (Vigna subterranea (L.)Verdc.) INDUCED WITH
SODIUM AZIDE USING SODIUM DODECYL SULPHATE POLYACRLYAMIDE GEL
ELECTROPHORESIS (SDS-PAGE)}



\author[1]{Juliet Naworu}%
\affil[1]{University of Ibadan Faculty of Science}%


\vspace{-1em}



  
  \date{June 15, 2024}


\begingroup
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\selectlanguage{english}
\begin{abstract}
This study investigated the mutagenic tolerance of eight Bambara
groundnut genotypes to Sodium azide (NaN3) using SDS-PAGE. The seeds of
six genotypes; TVSu-86, TVSu-91, TVSu-186, TVSu-235, TVSu-242, TVSu-350
were collected from IITA and two landraces from Abia and Enugu local
markets. The seeds were treated with five concentrations:
0.00\%(control), 0.01\%, 0.03\%, 0.05\% and 0.07\% of NaN3 after
pre-soaking for 6hrs in distilled water and sown in pots arranged in a
Complete Randomized Design with three replicates. There was reduction in
germination percentage and growth characters as concentrations of NaN3
increases. Early flowering was recorded at 37 days mutated with 0.07\%
of NaN3 compared to control which flowered late at 42 days. NaN3(0.07\%)
caused lethal effect on Abia and Enugu landraces. There was no
significant (P\textgreater{}0.05) difference in yield traits among
mutants and control. Mutant seeds significantly (P\textless{}0.05)
increased protein content (19.12\%) at 0.05\% of NaN3 compared to
control(18.5\%). The number of seeds(0.99), seed yield(0.89) and pod
yield(0.96) strongly correlated with seeds per pod (0.85). The SDS-PAGE
revealed the presence of polypeptide bands in mutants compared to
control. TVSu-235 and TVSu-350 genotypes had higher tolerance and yield
traits to 0.01\% concentration of NaN3, thus could be further improved
in subsequent breeding.%
\end{abstract}%



\sloppy


\textbf{INTRODUCTION}

Bambara groundnut (\emph{Vigna subterranea} (L.)Verdc,)\emph{,} member
of the family Fabaceae is an indigenous, underutilized African legume
crop mainly grown by subsistence farmers for food (Bamishaiye \emph{et
al} ., 2011). Bambara groundnut is the third most important leguminous
crop after Groundnut and Cowpea in terms of production and consumption
with which it shares its origin of genetic diversity (Aremu \emph{et
al.,} 2006). Genetic variation is fundamental to successful breeding
programs in vegetatively and sexually propagated plants (Olawuyi\emph{et
al} ., 2015; Agbolade \emph{et al} ., 2016). This variation which occurs
naturally or artificially by inducing with physical, biological or
chemical mutagens has attracted the interest of plant breeders for many
decades. Mutation breeding is an aspect of conventional plant breeding
which involves physical and chemical mutation in order to assess genetic
variation resulting to improved varieties with better characteristics
(Wongpiyasatid \emph{et al} ., 2000; Mensah and Obadoni, 2007;
Arulbalachandran \emph{et al} ., 2009; Olawuyi \emph{et al} ., 2016).

Among numerous techniques available for assessing the genetic
variability and relatedness among crop germplasm, seed storage protein
analysis represents a valid alternative and improved approach to
varietal identification (Mennella \emph{et al} ., 1999). Iqbal \emph{et
al} . (2005) reported that grain storage protein profiling based on
SDS-PAGE can be employed for various purposes such as; characterization
of germplasm, varietal identification, biosystematics analysis,
determination of phylogenetic relationship between different species and
generation of pertinent information to complement evaluation
(Ghafoor\emph{et al} ., 2002).

Sodium azide (NaN\textsubscript{3}) is one of the constituents of
bactericide, pesticide and industrial nitrogen gas generator that may be
lethal or beneficial in several plants such as barley, maize and some
other crop species (Ilbas \emph{et al} ., 2005; Mostafa, 2011).

Therefore, this study investigated the effects of sodium azide on
morphological and protein contents of Bambara groundnut using SDS-PAGE.

\textbf{MATERIALS AND METHODS}

Collection of seed samples, experimental location and experimental
design

Six (6) Seeds of Bambara Genotypes: TVSu-86, TVSu-91, TVSu-186,
TVSu-235, TVSu-242, TVSu-350 were collected from the germplasm unit of
the International Institute of Tropical Agriculture (IITA), Ibadan,
Nigeria and two (2) landraces were collected from Abia State and Enugu
State, Nigeria.

The \emph{in vitro} and screen house experiments were carried out at the
Genetics and Molecular Laboratory and Nursery farm of the Department of
Botany, University of Ibadan, Nigeria respectively. The design was laid
out in Complete Randomized Design (CRD) comprising of 6 genotypes, 2
landraces, 5 concentrations and 3 replicates.

Preparation of sodium azide (NaN\textsubscript{3}), pre-treatment of
seeds and in vitro screening

The seeds of Bambara were treated with concentrations of sodium azide
(0.01\%, 0.03\%, 0.05\% and 0.07\%) after pre-soaked in 100ml of
distilled for six(6) hours at room temperature (25\textsuperscript{o}C),
while untreated seeds (0.00\%) served as control according to the method
described by Asad \emph{et al.} (2014). Five seeds of both treated and
pre-soaked seeds were blotted with Whatman filter paper to remove excess
moisture for 20minutes and planted on Petri dishes lined with moist
filter paper. 2ml of distilled water was added daily to the petri dish
according to the method described by Asgharipour and Rafiei (2011).

Research design and Screen house experiment

10kg of sterilized top soil was measured using a weighing scale and put
into 120 perforated polythene bags. Three treated and untreated seeds of
each Bambara genotype were sown 3 inches deep inside polythene bag
filled with sterilized soil and spaced at a distance of 30cm in a
completely randomized design (CRD). Watering and other agronomic
practices were also carried out during growth and development of the
plant until maturity.

Determination of Quantitative characters and Protein contents

Observations on morphological traits of NaN\textsubscript{3} induced
mutants and controls were carried out and measured using measuring tape
and visual counting for a period of 14 weeks after planting following
the IPGRI, IITA and BAMNET (2000) Descriptors for Bambara groundnut. The
crude protein content was determined using micro Kjeldahl method as
described in AOAC (1996) and Sodium Dodecyl Sulphate - Polyacrylamide
Gel Electrophoresis (SDS-PAGE)was analysed using 12\% vertical slab
polyacrylamide gel for the separation of polypeptide bands according to
the procedure described by Weber and Osborn (1975).

\textbf{Statistical analysis}

Data were analyzed by analysis of variance (ANOVA) using SAS generalized
linear model (GLM) software and means with significant differences were
separated by Duncan multiple range test (DMRT) (P[?]0.05).The final
germination percentage (FGP) was obtained using Cokkizgin and Cokkizgin
(2010) method as stated below;

\begin{equation}
\text{\ FGP}=\frac{number\ of\ germinated\ seeds\ after\ 8\ days}{\text{total\ number\ of\ seeds\ planted}}\text{\ \ \ }\times\ \ 100\nonumber \\
 \end{equation}

\textbf{RESULTS AND DISCUSSION}

There was reduction in the germination percentage and growth characters
as the concentration of sodium azide increases. This could be as a
result of the lethal effect caused by the mutagen. Similar reductions
were also observed in maize in the reports made by Olawuyi and Okoli
(2017).

\textbf{Table 1. Mean square interaction of genotype, concentrations of
Sodium azide and growth stage on germination and growth characters of
Bambara groundnut}\selectlanguage{english}
\begin{longtable}[]{@{}llllllll@{}}
\toprule
Source of variation & Df & Germination percentage & Plant height & Leaf
length & Number of leaves & Number of branches & Leaf
width\tabularnewline
\midrule
\endhead
Concentration & 4 & 9.69\textsuperscript{ns} & 2121.02*** & 134.37*** &
42476.98*** & 5564.41*** & 12.95***\tabularnewline
Genotype & 7 & 385.48*** & 3321.67*** & 274.36*** & 143861.73*** &
17441.72*** & 25.47***\tabularnewline
Replicate & 2 & 2.32\textsuperscript{ns} & 60.89 \textsuperscript{ns} &
11.92 \textsuperscript{ns} & 5453.61 \textsuperscript{ns} & 562.16
\textsuperscript{ns} & 0.39\textsuperscript{ns}\tabularnewline
Week & 13 & 17146.10*** & 34.80*** & 2.07\textsuperscript{ns} &
91419.258*** & 10820.11*** & 2.28***\tabularnewline
Genotype \selectlanguage{ngerman}× concentration & 24 & 87.58\textsuperscript{ns} & 892.50*** &
52.29*** & 26745.69*** & 2853.22*** & 5.81***\tabularnewline
Concentration \selectlanguage{ngerman}× replicate & 8 & 74.82\textsuperscript{ns} & 120.31*** &
5.18*** & 25610.86*** & 3196.51*** & 0.71***\tabularnewline
Concentration \selectlanguage{ngerman}× week & 52 & 99.59*** & 30.14*** & 2.18*** & 1828.05*** &
222.47*** & 0.59***\tabularnewline
Genotype \selectlanguage{ngerman}× replicate & 14 & 73.01\textsuperscript{ns} & 66.85*** &
3.29*** & 3478.95*** & 412.41*** & 0.90***\tabularnewline
Genotype \selectlanguage{ngerman}× week & 91 & 385.60*** & 47.53*** & 3.61*** & 3786.25*** &
424.07*** & 0.45***\tabularnewline
Week \selectlanguage{ngerman}× replicate & 26 & 10.52\textsuperscript{ns} &
11.59\textsuperscript{ns} & 0.69\textsuperscript{ns} &
539.00\textsuperscript{ns} & 64.23\textsuperscript{ns} &
0.13\textsuperscript{ns}\tabularnewline
Genotype \selectlanguage{ngerman}× concentration × replicate & 47 & 45.91\textsuperscript{ns} &
117.29*** & 10.33*** & 13250.09*** & 1560.97*** & 0.89***\tabularnewline
Genotype \selectlanguage{ngerman}× concentration × week & 312 & 87.58*** & 32.89*** & 3.05*** &
2166.30** & 223.45*** & 0.53***\tabularnewline
Concentration \selectlanguage{ngerman}× week × replicate & 104 & 74.82*** &
15.29\textsuperscript{ns} & 1.52* & 1629.64*** & 214.31*** &
0.23\textsuperscript{ns}\tabularnewline
Genotype \selectlanguage{ngerman}× week × replicate & 182 & 73.01*** & 18.88* &
1.39\textsuperscript{ns} & 1094.67\textsuperscript{ns} &
127.02\textsuperscript{ns} & 0.23\textsuperscript{ns}\tabularnewline
Error & 611 & 611 & 562 & 562 & 560 & 560 & 533\tabularnewline
Corrected total & 1497 & 1497 & 1446 & 1446 & 1444 & 1444 &
1414\tabularnewline
\bottomrule
\end{longtable}

\emph{Note: * = significant at p\textless{}0.05, ** = highly significant
at p\textless{}0.01, *** = highly significant at p\textless{}0.001}

Early flowering was recorded in Bambara groundnut mutated with 0.07\% of
sodium azide compared to the control which flowered late. There was no
significant difference in yield traits among mutants and control, but
the mutants and control of landraces could not survive the environment,
therefore did not produce yield.

\textbf{Table 2. Mean square interactive effect of genotype,
concentration of sodium azide and growth stages on agronomic and yield
characters of Bambara groundnut}\selectlanguage{english}
\begin{longtable}[]{@{}llllllll@{}}
\toprule
Source of variation & df & Number of days to flowering & Seed yield per
plant (g) & Seeds per pod & Pod yield per plant (g) & Number of seeds
per plant & Number of pods per plant\tabularnewline
\midrule
\endhead
Concentration & 4 & 34.51*** & 0.35\textsuperscript{ns} &
0.01\textsuperscript{ns} & 1.21\textsuperscript{ns} &
2.22\textsuperscript{ns} & 2.19\textsuperscript{ns}\tabularnewline
Genotype & 7 & 135.99*** & 1.63*** & 0.02*** & 3.64*** & 7.78*** &
7.64***\tabularnewline
Replicate & 2 & 2.29\textsuperscript{ns} & 8.32 \textsuperscript{ns} &
0.32 \textsuperscript{ns} & 18.55 \textsuperscript{ns} & 46.02
\textsuperscript{ns} & 45.11 \textsuperscript{ns}\tabularnewline
Week & 13 & 10809.03*** & 8.55*** & 0.33*** & 19.40*** & 48.70*** &
47.78***\tabularnewline
Genotype \selectlanguage{ngerman}× concentration & 24 & 18.39*** & 0.43\textsuperscript{ns} &
0.004\textsuperscript{ns} & 0.52\textsuperscript{ns} &
1.06\textsuperscript{ns} & 1.04\textsuperscript{ns}\tabularnewline
Concentration \selectlanguage{ngerman}× replicate & 8 & 18.12*** & 0.22\textsuperscript{ns} &
0.01\textsuperscript{ns} & 0.68\textsuperscript{ns} &
1.23\textsuperscript{ns} & 1.23\textsuperscript{ns}\tabularnewline
Concentration \selectlanguage{ngerman}× week & 52 & 27.13*** & 0.33ns & 0.01*** & 1.22*** &
2.24** & 2.22***\tabularnewline
Genotype \selectlanguage{ngerman}× replicate & 14 & 3.34\textsuperscript{ns} & 1.73*** & 0.03***
& 3.99*** & 8.76*** & 8.61***\tabularnewline
Genotype \selectlanguage{ngerman}× week & 91 & 113.7* & 1.63*** & 0.03*** & 3.68*** & 7.87*** &
7.74***\tabularnewline
Week \selectlanguage{ngerman}× replicate & 26 & 1.71\textsuperscript{ns} & 5.50** & 0.21*** &
11.89*** & 30.42*** & 29.83***\tabularnewline
Genotype \selectlanguage{ngerman}× concentration × replicate & 47 & 3.44\textsuperscript{ns} &
0.46\textsuperscript{ns} & 0.01\textsuperscript{ns} &
0.60\textsuperscript{ns} & 1.22\textsuperscript{ns} &
1.19\textsuperscript{ns}\tabularnewline
Genotype \selectlanguage{ngerman}× concentration × week & 309 & 14.20*** &
0.43\textsuperscript{ns} & 0.004\textsuperscript{ns} &
0.52\textsuperscript{ns} & 1.06\textsuperscript{ns} &
1.04\textsuperscript{ns}\tabularnewline
Concentration \selectlanguage{ngerman}× week × replicate & 104 & 14.29*** &
0.21\textsuperscript{ns} & 0.05\textsuperscript{ns} &
0.68\textsuperscript{ns} & 1.24\textsuperscript{ns} &
1.23\textsuperscript{ns}\tabularnewline
Genotype \selectlanguage{ngerman}× week × replicate & 182 & 2.48\textsuperscript{ns} & 1.71*** &
0.03*** & 3.99*** & 8.76*** & 8.61***\tabularnewline
Error & 594 & 3.54 & 0.46 & 0.005 & 0.61 & 1.22 & 1.19\tabularnewline
Corrected total & 1477 & & & & & &\tabularnewline
\bottomrule
\end{longtable}

\emph{Note: * = significant at p\textless{}0.05, ** = highly significant
at p\textless{}0.01, *** = highly significant at p\textless{}0.001}

\textbf{Table 3. Genotypic effect on germination and growth response of
Bambara groundnut to sodium azide}\selectlanguage{english}
\begin{longtable}[]{@{}lllllll@{}}
\toprule
Genotype & Germination percentage (\%) & Plant length (cm) & Leaf length
(cm) & Number of leaves & Number of branches & Leaf width
(cm)\tabularnewline
\midrule
\endhead
TVSu 86 & 5.24\textsuperscript{a} & 21.51\textsuperscript{d} &
5.45\textsuperscript{d} & 124.09\textsuperscript{a} &
42.49\textsuperscript{a} & 1.93\textsuperscript{d}\tabularnewline
TVSu 91 & 2.54\textsuperscript{bc} & 12.80\textsuperscript{g} &
2.98\textsuperscript{f} & 57.74\textsuperscript{d} &
19.63\textsuperscript{e} & 1.16\textsuperscript{f}\tabularnewline
TVSu 186 & 4.77\textsuperscript{a} & 26.21\textsuperscript{a} &
7.11\textsuperscript{a} & 117.78\textsuperscript{ab} &
41.03\textsuperscript{ab} & 2.26\textsuperscript{b}\tabularnewline
TVSu 235 & 4.92\textsuperscript{a} & 25.08\textsuperscript{b} &
6.07\textsuperscript{c} & 118.28\textsuperscript{ab} &
40.20\textsuperscript{ab} & 2.44\textsuperscript{a}\tabularnewline
TVSu 242 & 2.38\textsuperscript{bc} & 23.00\textsuperscript{c} &
5.95\textsuperscript{c} & 73.46\textsuperscript{c} &
26.02\textsuperscript{c} & 2.13\textsuperscript{c}\tabularnewline
TVSu 350 & 3.81\textsuperscript{ab} & 25.53\textsuperscript{ab} &
6.69\textsuperscript{b} & 110.19\textsuperscript{b} &
38.65\textsuperscript{b} & 2.43\textsuperscript{a}\tabularnewline
Abia & 1.59\textsuperscript{c} & 18.76\textsuperscript{f} &
4.73\textsuperscript{e} & 68.52\textsuperscript{c} &
23.28\textsuperscript{d} & 2.19\textsuperscript{bc}\tabularnewline
Enugu & 4.76\textsuperscript{a} & 20.00\textsuperscript{e} &
4.92\textsuperscript{e} & 43.64\textsuperscript{e} &
15.05\textsuperscript{f} & 1.69\textsuperscript{e}\tabularnewline
\bottomrule
\end{longtable}

\emph{Means with different letter within a column differ significantly
at p\textless{}0.05}

\textbf{Table 4. Effect of sodium azide concentrations on growth of
Bambara groundnut}\selectlanguage{english}
\begin{longtable}[]{@{}lllllll@{}}
\toprule
Sodium azide concentrations (\%) & Germination (\%) & Plant length (cm)
& Leaf length (cm) & Number of leaves & Number of branches & Leaf width
(cm)\tabularnewline
\midrule
\endhead
0.00 (Control) & 4.37\textsuperscript{a} & 25.10\textsuperscript{a} &
6.33\textsuperscript{a} & 108.46\textsuperscript{a} &
37.51\textsuperscript{a} & 2.28\textsuperscript{a}\tabularnewline
0.01 & 3.74\textsuperscript{ab} & 22.41\textsuperscript{b} &
5.84\textsuperscript{b} & 93.12\textsuperscript{b} &
32.77\textsuperscript{b} & 2.29\textsuperscript{a}\tabularnewline
0.03 & 3.18\textsuperscript{b} & 20.12\textsuperscript{c} &
5.18\textsuperscript{c} & 83.35\textsuperscript{c} &
28.30\textsuperscript{c} & 1.94\textsuperscript{c}\tabularnewline
0.05 & 3.86\textsuperscript{ab} & 23.03\textsuperscript{b} &
5.83\textsuperscript{b} & 95.48\textsuperscript{b} &
32.59\textsuperscript{b} & 2.09\textsuperscript{b}\tabularnewline
0.07 & 2.98\textsuperscript{b} & 18.15\textsuperscript{d} &
4.58\textsuperscript{d} & 93.38\textsuperscript{b} &
32.36\textsuperscript{b} & 1.69\textsuperscript{d}\tabularnewline
\bottomrule
\end{longtable}

\emph{Means with different letter within a column differ significantly
at p\textless{}0.05}

Mutant seeds had significant increase in protein content. The
concentration of 0.05\% sodium azide had the highest protein content
(19.12\%), while control had (18.5\%). The increase in protein content
at higher concentration of sodium azide could be due to increase in
auxin level. Similar results were observed in French bean by
Mahamune\emph{et al} . (2017).

\textbf{Table 5. Genotypic effects on protein contents, agronomic and
yield of Bambara groundnut induced with sodium azide}\selectlanguage{english}
\begin{longtable}[]{@{}llllllll@{}}
\toprule
Genotype & Number of days to flowering & Seed yield per plant (g) &
Seeds per pod & Pod yield per plant (g) & Number of seeds per plant &
Number of pods per plant & Protein contents (\%)\tabularnewline
\midrule
\endhead
TVSu 86 & 2.86\textsuperscript{cd} & 0.17\textsuperscript{ab} &
0.03\textsuperscript{a} & 0.22\textsuperscript{abc} &
0.32\textsuperscript{ab} & 0.31\textsuperscript{ab} &
18.50\textsuperscript{e}\tabularnewline
TVSu 91 & 2.04\textsuperscript{e} & 0.02\textsuperscript{b} &
0.01\textsuperscript{bc} & 0.03\textsuperscript{dc} &
0.06\textsuperscript{bc} & 0.05\textsuperscript{bc} &
19.15\textsuperscript{b}\tabularnewline
TVSu 186 & 3.44\textsuperscript{b} & 0.05\textsuperscript{b} &
0.02\textsuperscript{ab} & 0.08\textsuperscript{bcd} &
0.14\textsuperscript{bc} & 0.14\textsuperscript{bc} &
18.85\textsuperscript{d}\tabularnewline
TVSu 235 & 2.49\textsuperscript{d} & 0.25\textsuperscript{a} &
0.03\textsuperscript{a} & 0.37\textsuperscript{a} &
0.50\textsuperscript{a} & 0.49\textsuperscript{a} &
19.38\textsuperscript{a}\tabularnewline
TVSu 242 & 3.59\textsuperscript{b} & 0.02\textsuperscript{b} &
0.01\textsuperscript{bc} & 0.04\textsuperscript{dc} &
0.08\textsuperscript{bc} & 0.08\textsuperscript{bc} &
19.00\textsuperscript{c}\tabularnewline
TVSu 350 & 3.21\textsuperscript{bc} & 0.14\textsuperscript{ab} &
0.02\textsuperscript{ab} & 0.25\textsuperscript{ab} &
0.46\textsuperscript{a} & 0.46\textsuperscript{a} &
18.80\textsuperscript{d}\tabularnewline
Abia & 1.89\textsuperscript{e} & 0.00\textsuperscript{b} &
0.00\textsuperscript{c} & 0.02\textsuperscript{dc} &
0.00\textsuperscript{c} & 0.02\textsuperscript{d} & -\tabularnewline
Enugu & 6.76\textsuperscript{a} & 0.00\textsuperscript{b} &
0.00\textsuperscript{c} & 0.00\textsuperscript{d} &
0.00\textsuperscript{c} & 0.00\textsuperscript{c} & -\tabularnewline
\bottomrule
\end{longtable}

\emph{Means with different letter within a column differ significantly
at p\textless{}0.05}

\textbf{Table 6. Effect of sodium azide concentrations on protein
contents, agronomic and yield characters of Bambara groundnut}\selectlanguage{english}
\begin{longtable}[]{@{}llllllll@{}}
\toprule
Sodium azide concentrations & Number of days to flowering & Seed yield
per plant (g) & Seeds per pod & Pod yield per plant (g) & Number of
seeds per plant & Number of pods per plant & Protein contents
(\%)\tabularnewline
\midrule
\endhead
0.00\% (Control) & 3.49\textsuperscript{a} & 0.12\textsuperscript{a} &
0.02\textsuperscript{a} & 0.22\textsuperscript{a} &
0.31\textsuperscript{a} & 0.30\textsuperscript{a} &
18.75\textsuperscript{c}\tabularnewline
0.01\% & 3.04\textsuperscript{b} & 0.07\textsuperscript{a} &
0.02\textsuperscript{a} & 0.11\textsuperscript{a} &
0.28\textsuperscript{a} & 0.20\textsuperscript{a} &
18.76\textsuperscript{c}\tabularnewline
0.03\% & 2.60\textsuperscript{c} & 0.05\textsuperscript{a} &
0.01\textsuperscript{a} & 0.20\textsuperscript{a} &
0.20\textsuperscript{a} & 0.14\textsuperscript{a} &
18.95\textsuperscript{b}\tabularnewline
0.05\% & 3.15\textsuperscript{b} & 0.10\textsuperscript{a} &
0.02\textsuperscript{a} & 0.16\textsuperscript{a} &
0.27\textsuperscript{a} & 0.27\textsuperscript{a} &
19.12\textsuperscript{a}\tabularnewline
0.07\% & 2.08\textsuperscript{d} & 0.10\textsuperscript{a} &
0.00\textsuperscript{a} & 0.11\textsuperscript{a} &
0.18\textsuperscript{a} & 0.18\textsuperscript{a} &
19.06\textsuperscript{a}\tabularnewline
\bottomrule
\end{longtable}

\emph{Means with different letter within a column differ significantly
at p\textless{}0.05}

\textbf{Table 7. Correlation coefficient of growth, agronomic and yield
characters of Bambara groundnut induced with sodium azide.}\selectlanguage{english}
\begin{longtable}[]{@{}lllllllllllllllll@{}}
\toprule
& G \% & NB & PH & NL & LL & LW & DF & PP & NS & PY & SY & S/P & G & C &
W & R\tabularnewline
\midrule
\endhead
Germination \% & & & & & & & & & & & & & & & &\tabularnewline
Number of branches & -0.24 & & & & & & & & & & & & & & &\tabularnewline
Plant height & 0.03 & 0.51* & & & & & & & & & & & & & &\tabularnewline
Number of leaves & -0.23 & 0.98** & 0.50* & & & & & & & & & & & &
&\tabularnewline
Leaf length & 0.04 & 0.48 & 0.85** & 0.47 & & & & & & & & & & &
&\tabularnewline
Leaf width & -0.05 & 0.45 & 0.74** & 0.45 & 0.71** & & & & & & & & & &
&\tabularnewline
Days to flowering & -0.05 & 0.10 & 0.04 & 0.07 & 0.03 & 0.05 & & & & & &
& & & &\tabularnewline
Pods per plant & -0.024 & 0.09 & 0.03 & 0.07 & 0.03 & 0.04 & 0.35 & & &
& & & & & &\tabularnewline
Number of seeds & -0.02 & 0.09 & 0.03 & 0.07 & 0.03 & 0.04 & 0.35 &
0.99** & & & & & & & &\tabularnewline
Pod yield (g) & -0.02 & 0.08 & 0.02 & 0.07 & 0.03 & 0.04 & 0.33 & 0.96**
& 0.96** & & & & & & &\tabularnewline
Seed yield (g) & -0.02 & 0.07 & 0.02 & 0.06 & 0.02 & 0.03 & 0.29 &
0.89** & 0.90** & 0.94** & & & & & &\tabularnewline
Seeds per pod & -0.03 & 0.08 & 0.03 & 0.06 & 0.03 & 0.05 & 0.38 & 0.85**
& 0.85** & 0.86** & 0.84** & & & & &\tabularnewline
Genotype & -0.04 & -0.12 & 0.18 & -0.12 & 0.18 & -0.12 & 0.21 & 0.25 &
0.01 & -0.01 & -0.02 & -0.04 & & & &\tabularnewline
Concentration & -0.02 & -0.06 & -0.23 & -0.06 & -0.21 & -0.22 & 0.02 &
-0.02 & -0.02 & -0.02 & -0.01 & -0.02 & -0.09 & & &\tabularnewline
Week & -0.37 & 0.51* & 0.11 & 0.49 & 0.05 & 0.16 & 0.40 & 0.18 & 0.18 &
0.17 & 0.16 & 0.20 & -0.02 & -0.03 & &\tabularnewline
Replicates & -0.00 & 0.01 & 0.04 & 0.02 & 0.01 & 0.01 & 0.00 & -0.12 &
-0.12 & -0.12 & -0.11 & -0.14 & 0.01 & 0.03 & -0.00 &\tabularnewline
\bottomrule
\end{longtable}

\emph{Note: * = related (correlated) at p\textless{}0.05, ** = strongly
related at p\textless{}0.01. G\% = Germination percentage, NB = Number
of branches, PH = Plant height, NL = Number of leaves, LL = Leaf length,
LW = Leaf width, DF = Days to flowering, PP = Pods per plant, NS =
Number of seeds, PY = Pod yield, SY = Seed yield, S/P = Seeds per pod, G
= Genotype, C = Concentration, W = Week and R = Replicate}

The SDS-PAGE analysis revealed the presence of polypeptide bands in
mutants which could be due to the damaging effect of the mutagen on
different genes responsible for the polypeptide subunits. Similar
occurrence had also been reported in Cowpea by Odeigah \emph{et
al.}(1998) and Mahamune \emph{et al} . (2017).

The consumption of any legumes may result in flatulence and constipation
which could be attributed to higher concentration of sodium azide in
pesticide used in controlling pest damaging field crops (Ilbas \emph{et
al.,} 2005).

\textbf{CONCLUSION}

The concentration of 0.07\% Sodium azide mutagen reduced germination,
growth and yield but increased the protein content of Bambara groundnut
compared with control.TVSu-235 and TVSu-350 genotypes had higher
tolerance and yield response to sodium azide concentrations therefore;
they could be further utilised in crop improvement. The growth and yield
traits should also be encouraged in mutagenic breeding of other
leguminous crops.

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application of different concentrations of NaN\textsubscript{3} for
different times on the morphological and cytogenetic characteristics of
barley (\emph{Hordeum vulgare} L.) seedlings. \emph{Journal of
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SDS-PAGE markers and Ascochyta blight in chickpea. \emph{Pakistan
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Olawuyi, O. J., Bello, O.B., Ntube, C.V., \& Akanmu, A.O. (2015).
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Olawuyi, O. J., \& Okoli, S.O. (2017). Genetic variability on tolerance
of maize (\emph{Zea mays} L\emph{.)} genotypes induced with sodium azide
mutagen. \emph{Molecular Plant Breeding,} 8(3): 27-37.

Odeigah, P.G.C. \& Osanyinpeju, A.O. (1998). Induced mutation in
cowpea\emph{(Vigna unguiculata} ). \emph{J. Genet Breeding,} 51:
126-132.

Weber, K. \& M. Osborn, 1975. Proteins and Sodium dodecyl sulphate.
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Promcham, W. (2000). Induced mutations in Mungbean breeding: Regional
yield trial of mungbean mutant lines. \emph{Kasetsart J. Nat. Sci.
Res.,} 34: 443-449.

\textbf{Table 1. Mean square interaction of genotype, concentrations of
Sodium azide and growth stage on germination and growth characters of
Bambara groundnut}\selectlanguage{english}
\begin{longtable}[]{@{}llllllll@{}}
\toprule
Source of variation & Df & Germination percentage & Plant height & Leaf
length & Number of leaves & Number of branches & Leaf
width\tabularnewline
\midrule
\endhead
Concentration & 4 & 9.69\textsuperscript{ns} & 2121.02*** & 134.37*** &
42476.98*** & 5564.41*** & 12.95***\tabularnewline
Genotype & 7 & 385.48*** & 3321.67*** & 274.36*** & 143861.73*** &
17441.72*** & 25.47***\tabularnewline
Replicate & 2 & 2.32\textsuperscript{ns} & 60.89 \textsuperscript{ns} &
11.92 \textsuperscript{ns} & 5453.61 \textsuperscript{ns} & 562.16
\textsuperscript{ns} & 0.39\textsuperscript{ns}\tabularnewline
Week & 13 & 17146.10*** & 34.80*** & 2.07\textsuperscript{ns} &
91419.258*** & 10820.11*** & 2.28***\tabularnewline
Genotype \selectlanguage{ngerman}× concentration & 24 & 87.58\textsuperscript{ns} & 892.50*** &
52.29*** & 26745.69*** & 2853.22*** & 5.81***\tabularnewline
Concentration \selectlanguage{ngerman}× replicate & 8 & 74.82\textsuperscript{ns} & 120.31*** &
5.18*** & 25610.86*** & 3196.51*** & 0.71***\tabularnewline
Concentration \selectlanguage{ngerman}× week & 52 & 99.59*** & 30.14*** & 2.18*** & 1828.05*** &
222.47*** & 0.59***\tabularnewline
Genotype \selectlanguage{ngerman}× replicate & 14 & 73.01\textsuperscript{ns} & 66.85*** &
3.29*** & 3478.95*** & 412.41*** & 0.90***\tabularnewline
Genotype \selectlanguage{ngerman}× week & 91 & 385.60*** & 47.53*** & 3.61*** & 3786.25*** &
424.07*** & 0.45***\tabularnewline
Week \selectlanguage{ngerman}× replicate & 26 & 10.52\textsuperscript{ns} &
11.59\textsuperscript{ns} & 0.69\textsuperscript{ns} &
539.00\textsuperscript{ns} & 64.23\textsuperscript{ns} &
0.13\textsuperscript{ns}\tabularnewline
Genotype \selectlanguage{ngerman}× concentration × replicate & 47 & 45.91\textsuperscript{ns} &
117.29*** & 10.33*** & 13250.09*** & 1560.97*** & 0.89***\tabularnewline
Genotype \selectlanguage{ngerman}× concentration × week & 312 & 87.58*** & 32.89*** & 3.05*** &
2166.30** & 223.45*** & 0.53***\tabularnewline
Concentration \selectlanguage{ngerman}× week × replicate & 104 & 74.82*** &
15.29\textsuperscript{ns} & 1.52* & 1629.64*** & 214.31*** &
0.23\textsuperscript{ns}\tabularnewline
Genotype \selectlanguage{ngerman}× week × replicate & 182 & 73.01*** & 18.88* &
1.39\textsuperscript{ns} & 1094.67\textsuperscript{ns} &
127.02\textsuperscript{ns} & 0.23\textsuperscript{ns}\tabularnewline
Error & 611 & 611 & 562 & 562 & 560 & 560 & 533\tabularnewline
Corrected total & 1497 & 1497 & 1446 & 1446 & 1444 & 1444 &
1414\tabularnewline
\bottomrule
\end{longtable}

\emph{Note: * = significant at p\textless{}0.05, ** = highly significant
at p\textless{}0.01, *** = highly significant at p\textless{}0.001}

\textbf{Table 2. Mean square interactive effect of genotype,
concentration of sodium azide and growth stages on agronomic and yield
characters of Bambara groundnut}\selectlanguage{english}
\begin{longtable}[]{@{}llllllll@{}}
\toprule
Source of variation & df & Number of days to flowering & Seed yield per
plant (g) & Seeds per pod & Pod yield per plant (g) & Number of seeds
per plant & Number of pods per plant\tabularnewline
\midrule
\endhead
Concentration & 4 & 34.51*** & 0.35\textsuperscript{ns} &
0.01\textsuperscript{ns} & 1.21\textsuperscript{ns} &
2.22\textsuperscript{ns} & 2.19\textsuperscript{ns}\tabularnewline
Genotype & 7 & 135.99*** & 1.63*** & 0.02*** & 3.64*** & 7.78*** &
7.64***\tabularnewline
Replicate & 2 & 2.29\textsuperscript{ns} & 8.32 \textsuperscript{ns} &
0.32 \textsuperscript{ns} & 18.55 \textsuperscript{ns} & 46.02
\textsuperscript{ns} & 45.11 \textsuperscript{ns}\tabularnewline
Week & 13 & 10809.03*** & 8.55*** & 0.33*** & 19.40*** & 48.70*** &
47.78***\tabularnewline
Genotype \selectlanguage{ngerman}× concentration & 24 & 18.39*** & 0.43\textsuperscript{ns} &
0.004\textsuperscript{ns} & 0.52\textsuperscript{ns} &
1.06\textsuperscript{ns} & 1.04\textsuperscript{ns}\tabularnewline
Concentration \selectlanguage{ngerman}× replicate & 8 & 18.12*** & 0.22\textsuperscript{ns} &
0.01\textsuperscript{ns} & 0.68\textsuperscript{ns} &
1.23\textsuperscript{ns} & 1.23\textsuperscript{ns}\tabularnewline
Concentration \selectlanguage{ngerman}× week & 52 & 27.13*** & 0.33ns & 0.01*** & 1.22*** &
2.24** & 2.22***\tabularnewline
Genotype \selectlanguage{ngerman}× replicate & 14 & 3.34\textsuperscript{ns} & 1.73*** & 0.03***
& 3.99*** & 8.76*** & 8.61***\tabularnewline
Genotype \selectlanguage{ngerman}× week & 91 & 113.7* & 1.63*** & 0.03*** & 3.68*** & 7.87*** &
7.74***\tabularnewline
Week \selectlanguage{ngerman}× replicate & 26 & 1.71\textsuperscript{ns} & 5.50** & 0.21*** &
11.89*** & 30.42*** & 29.83***\tabularnewline
Genotype \selectlanguage{ngerman}× concentration × replicate & 47 & 3.44\textsuperscript{ns} &
0.46\textsuperscript{ns} & 0.01\textsuperscript{ns} &
0.60\textsuperscript{ns} & 1.22\textsuperscript{ns} &
1.19\textsuperscript{ns}\tabularnewline
Genotype \selectlanguage{ngerman}× concentration × week & 309 & 14.20*** &
0.43\textsuperscript{ns} & 0.004\textsuperscript{ns} &
0.52\textsuperscript{ns} & 1.06\textsuperscript{ns} &
1.04\textsuperscript{ns}\tabularnewline
Concentration \selectlanguage{ngerman}× week × replicate & 104 & 14.29*** &
0.21\textsuperscript{ns} & 0.05\textsuperscript{ns} &
0.68\textsuperscript{ns} & 1.24\textsuperscript{ns} &
1.23\textsuperscript{ns}\tabularnewline
Genotype \selectlanguage{ngerman}× week × replicate & 182 & 2.48\textsuperscript{ns} & 1.71*** &
0.03*** & 3.99*** & 8.76*** & 8.61***\tabularnewline
Error & 594 & 3.54 & 0.46 & 0.005 & 0.61 & 1.22 & 1.19\tabularnewline
Corrected total & 1477 & & & & & &\tabularnewline
\bottomrule
\end{longtable}

\emph{Note: * = significant at p\textless{}0.05, ** = highly significant
at p\textless{}0.01, *** = highly significant at p\textless{}0.001}

\textbf{Table 3. Genotypic effect on germination and growth response of
Bambara groundnut to sodium azide}\selectlanguage{english}
\begin{longtable}[]{@{}lllllll@{}}
\toprule
Genotype & Germination percentage (\%) & Plant length (cm) & Leaf length
(cm) & Number of leaves & Number of branches & Leaf width
(cm)\tabularnewline
\midrule
\endhead
TVSu 86 & 5.24\textsuperscript{a} & 21.51\textsuperscript{d} &
5.45\textsuperscript{d} & 124.09\textsuperscript{a} &
42.49\textsuperscript{a} & 1.93\textsuperscript{d}\tabularnewline
TVSu 91 & 2.54\textsuperscript{bc} & 12.80\textsuperscript{g} &
2.98\textsuperscript{f} & 57.74\textsuperscript{d} &
19.63\textsuperscript{e} & 1.16\textsuperscript{f}\tabularnewline
TVSu 186 & 4.77\textsuperscript{a} & 26.21\textsuperscript{a} &
7.11\textsuperscript{a} & 117.78\textsuperscript{ab} &
41.03\textsuperscript{ab} & 2.26\textsuperscript{b}\tabularnewline
TVSu 235 & 4.92\textsuperscript{a} & 25.08\textsuperscript{b} &
6.07\textsuperscript{c} & 118.28\textsuperscript{ab} &
40.20\textsuperscript{ab} & 2.44\textsuperscript{a}\tabularnewline
TVSu 242 & 2.38\textsuperscript{bc} & 23.00\textsuperscript{c} &
5.95\textsuperscript{c} & 73.46\textsuperscript{c} &
26.02\textsuperscript{c} & 2.13\textsuperscript{c}\tabularnewline
TVSu 350 & 3.81\textsuperscript{ab} & 25.53\textsuperscript{ab} &
6.69\textsuperscript{b} & 110.19\textsuperscript{b} &
38.65\textsuperscript{b} & 2.43\textsuperscript{a}\tabularnewline
Abia & 1.59\textsuperscript{c} & 18.76\textsuperscript{f} &
4.73\textsuperscript{e} & 68.52\textsuperscript{c} &
23.28\textsuperscript{d} & 2.19\textsuperscript{bc}\tabularnewline
Enugu & 4.76\textsuperscript{a} & 20.00\textsuperscript{e} &
4.92\textsuperscript{e} & 43.64\textsuperscript{e} &
15.05\textsuperscript{f} & 1.69\textsuperscript{e}\tabularnewline
\bottomrule
\end{longtable}

\emph{Means with different letter within a column differ significantly
at p\textless{}0.05}

\textbf{Table 4. Effect of sodium azide concentrations on growth of
Bambara groundnut}\selectlanguage{english}
\begin{longtable}[]{@{}lllllll@{}}
\toprule
Sodium azide concentrations (\%) & Germination (\%) & Plant length (cm)
& Leaf length (cm) & Number of leaves & Number of branches & Leaf width
(cm)\tabularnewline
\midrule
\endhead
0.00 (Control) & 4.37\textsuperscript{a} & 25.10\textsuperscript{a} &
6.33\textsuperscript{a} & 108.46\textsuperscript{a} &
37.51\textsuperscript{a} & 2.28\textsuperscript{a}\tabularnewline
0.01 & 3.74\textsuperscript{ab} & 22.41\textsuperscript{b} &
5.84\textsuperscript{b} & 93.12\textsuperscript{b} &
32.77\textsuperscript{b} & 2.29\textsuperscript{a}\tabularnewline
0.03 & 3.18\textsuperscript{b} & 20.12\textsuperscript{c} &
5.18\textsuperscript{c} & 83.35\textsuperscript{c} &
28.30\textsuperscript{c} & 1.94\textsuperscript{c}\tabularnewline
0.05 & 3.86\textsuperscript{ab} & 23.03\textsuperscript{b} &
5.83\textsuperscript{b} & 95.48\textsuperscript{b} &
32.59\textsuperscript{b} & 2.09\textsuperscript{b}\tabularnewline
0.07 & 2.98\textsuperscript{b} & 18.15\textsuperscript{d} &
4.58\textsuperscript{d} & 93.38\textsuperscript{b} &
32.36\textsuperscript{b} & 1.69\textsuperscript{d}\tabularnewline
\bottomrule
\end{longtable}

\emph{Means with different letter within a column differ significantly
at p\textless{}0.05}

\textbf{Table 5. Genotypic effects on protein contents, agronomic and
yield of Bambara groundnut induced with sodium azide}\selectlanguage{english}
\begin{longtable}[]{@{}llllllll@{}}
\toprule
Genotype & Number of days to flowering & Seed yield per plant (g) &
Seeds per pod & Pod yield per plant (g) & Number of seeds per plant &
Number of pods per plant & Protein contents (\%)\tabularnewline
\midrule
\endhead
TVSu 86 & 2.86\textsuperscript{cd} & 0.17\textsuperscript{ab} &
0.03\textsuperscript{a} & 0.22\textsuperscript{abc} &
0.32\textsuperscript{ab} & 0.31\textsuperscript{ab} &
18.50\textsuperscript{e}\tabularnewline
TVSu 91 & 2.04\textsuperscript{e} & 0.02\textsuperscript{b} &
0.01\textsuperscript{bc} & 0.03\textsuperscript{dc} &
0.06\textsuperscript{bc} & 0.05\textsuperscript{bc} &
19.15\textsuperscript{b}\tabularnewline
TVSu 186 & 3.44\textsuperscript{b} & 0.05\textsuperscript{b} &
0.02\textsuperscript{ab} & 0.08\textsuperscript{bcd} &
0.14\textsuperscript{bc} & 0.14\textsuperscript{bc} &
18.85\textsuperscript{d}\tabularnewline
TVSu 235 & 2.49\textsuperscript{d} & 0.25\textsuperscript{a} &
0.03\textsuperscript{a} & 0.37\textsuperscript{a} &
0.50\textsuperscript{a} & 0.49\textsuperscript{a} &
19.38\textsuperscript{a}\tabularnewline
TVSu 242 & 3.59\textsuperscript{b} & 0.02\textsuperscript{b} &
0.01\textsuperscript{bc} & 0.04\textsuperscript{dc} &
0.08\textsuperscript{bc} & 0.08\textsuperscript{bc} &
19.00\textsuperscript{c}\tabularnewline
TVSu 350 & 3.21\textsuperscript{bc} & 0.14\textsuperscript{ab} &
0.02\textsuperscript{ab} & 0.25\textsuperscript{ab} &
0.46\textsuperscript{a} & 0.46\textsuperscript{a} &
18.80\textsuperscript{d}\tabularnewline
Abia & 1.89\textsuperscript{e} & 0.00\textsuperscript{b} &
0.00\textsuperscript{c} & 0.02\textsuperscript{dc} &
0.00\textsuperscript{c} & 0.02\textsuperscript{d} & -\tabularnewline
Enugu & 6.76\textsuperscript{a} & 0.00\textsuperscript{b} &
0.00\textsuperscript{c} & 0.00\textsuperscript{d} &
0.00\textsuperscript{c} & 0.00\textsuperscript{c} & -\tabularnewline
\bottomrule
\end{longtable}

\emph{Means with different letter within a column differ significantly
at p\textless{}0.05}

\textbf{Table 6. Effect of sodium azide concentrations on protein
contents, agronomic and yield characters of Bambara groundnut}\selectlanguage{english}
\begin{longtable}[]{@{}llllllll@{}}
\toprule
Sodium azide concentrations & Number of days to flowering & Seed yield
per plant (g) & Seeds per pod & Pod yield per plant (g) & Number of
seeds per plant & Number of pods per plant & Protein contents
(\%)\tabularnewline
\midrule
\endhead
0.00\% (Control) & 3.49\textsuperscript{a} & 0.12\textsuperscript{a} &
0.02\textsuperscript{a} & 0.22\textsuperscript{a} &
0.31\textsuperscript{a} & 0.30\textsuperscript{a} &
18.75\textsuperscript{c}\tabularnewline
0.01\% & 3.04\textsuperscript{b} & 0.07\textsuperscript{a} &
0.02\textsuperscript{a} & 0.11\textsuperscript{a} &
0.28\textsuperscript{a} & 0.20\textsuperscript{a} &
18.76\textsuperscript{c}\tabularnewline
0.03\% & 2.60\textsuperscript{c} & 0.05\textsuperscript{a} &
0.01\textsuperscript{a} & 0.20\textsuperscript{a} &
0.20\textsuperscript{a} & 0.14\textsuperscript{a} &
18.95\textsuperscript{b}\tabularnewline
0.05\% & 3.15\textsuperscript{b} & 0.10\textsuperscript{a} &
0.02\textsuperscript{a} & 0.16\textsuperscript{a} &
0.27\textsuperscript{a} & 0.27\textsuperscript{a} &
19.12\textsuperscript{a}\tabularnewline
0.07\% & 2.08\textsuperscript{d} & 0.10\textsuperscript{a} &
0.00\textsuperscript{a} & 0.11\textsuperscript{a} &
0.18\textsuperscript{a} & 0.18\textsuperscript{a} &
19.06\textsuperscript{a}\tabularnewline
\bottomrule
\end{longtable}

\emph{Means with different letter within a column differ significantly
at p\textless{}0.05}

\textbf{Table 7. Correlation coefficient of growth, agronomic and yield
characters of Bambara groundnut induced with sodium azide.}\selectlanguage{english}
\begin{longtable}[]{@{}lllllllllllllllll@{}}
\toprule
& G \% & NB & PH & NL & LL & LW & DF & PP & NS & PY & SY & S/P & G & C &
W & R\tabularnewline
\midrule
\endhead
Germination \% & & & & & & & & & & & & & & & &\tabularnewline
Number of branches & -0.24 & & & & & & & & & & & & & & &\tabularnewline
Plant height & 0.03 & 0.51* & & & & & & & & & & & & & &\tabularnewline
Number of leaves & -0.23 & 0.98** & 0.50* & & & & & & & & & & & &
&\tabularnewline
Leaf length & 0.04 & 0.48 & 0.85** & 0.47 & & & & & & & & & & &
&\tabularnewline
Leaf width & -0.05 & 0.45 & 0.74** & 0.45 & 0.71** & & & & & & & & & &
&\tabularnewline
Days to flowering & -0.05 & 0.10 & 0.04 & 0.07 & 0.03 & 0.05 & & & & & &
& & & &\tabularnewline
Pods per plant & -0.024 & 0.09 & 0.03 & 0.07 & 0.03 & 0.04 & 0.35 & & &
& & & & & &\tabularnewline
Number of seeds & -0.02 & 0.09 & 0.03 & 0.07 & 0.03 & 0.04 & 0.35 &
0.99** & & & & & & & &\tabularnewline
Pod yield (g) & -0.02 & 0.08 & 0.02 & 0.07 & 0.03 & 0.04 & 0.33 & 0.96**
& 0.96** & & & & & & &\tabularnewline
Seed yield (g) & -0.02 & 0.07 & 0.02 & 0.06 & 0.02 & 0.03 & 0.29 &
0.89** & 0.90** & 0.94** & & & & & &\tabularnewline
Seeds per pod & -0.03 & 0.08 & 0.03 & 0.06 & 0.03 & 0.05 & 0.38 & 0.85**
& 0.85** & 0.86** & 0.84** & & & & &\tabularnewline
Genotype & -0.04 & -0.12 & 0.18 & -0.12 & 0.18 & -0.12 & 0.21 & 0.25 &
0.01 & -0.01 & -0.02 & -0.04 & & & &\tabularnewline
Concentration & -0.02 & -0.06 & -0.23 & -0.06 & -0.21 & -0.22 & 0.02 &
-0.02 & -0.02 & -0.02 & -0.01 & -0.02 & -0.09 & & &\tabularnewline
Week & -0.37 & 0.51* & 0.11 & 0.49 & 0.05 & 0.16 & 0.40 & 0.18 & 0.18 &
0.17 & 0.16 & 0.20 & -0.02 & -0.03 & &\tabularnewline
Replicates & -0.00 & 0.01 & 0.04 & 0.02 & 0.01 & 0.01 & 0.00 & -0.12 &
-0.12 & -0.12 & -0.11 & -0.14 & 0.01 & 0.03 & -0.00 &\tabularnewline
\bottomrule
\end{longtable}

\emph{Note: * = related (correlated) at p\textless{}0.05, ** = strongly
related at p\textless{}0.01. G\% = Germination percentage, NB = Number
of branches, PH = Plant height, NL = Number of leaves, LL = Leaf length,
LW = Leaf width, DF = Days to flowering, PP = Pods per plant, NS =
Number of seeds, PY = Pod yield, SY = Seed yield, S/P = Seeds per pod, G
= Genotype, C = Concentration, W = Week and R = Replicate}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/image1/image1}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/image2/image2}

\end{center}
\end{figure}

\textbf{Plate 1a.} TVSu-186 (0.07\%) induced with \textbf{Plate 1b} .
TVSu-186 (0.00\%) control

Sodium azide showing purple seedling showing normal colour\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/image3/image3}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/image4/image4}

\end{center}
\end{figure}

\textbf{Plate 2a.} Abia landraces induced with \textbf{Plate 2b}
.TVSu-91 induced with 0.05\%

0.05\% Sodium azide showed creeping Sodium azide had two different
leaves shape

stem and yellow colour. in a single plant stand.\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/image5/image5}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/image6/image6}

\end{center}
\end{figure}

\textbf{Plate 3.} Abia landrace induced with 0.01\% \textbf{Plate
4.}TVSu-86 (0.07\%) produced mutants

Sodium azide showed crinkled (rough and with two seeds per pod which was
not

twisting) leaves. observed in other concentrations.\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/image7/image7}

\end{center}
\end{figure}

\textbf{Plate 5} . SDS-polyacrylamide gel electrophoregram of seed
protein profile.

\selectlanguage{english}
\FloatBarrier
\end{document}

