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International Journal of Plant Breeding and Crop Science

Vol. 6(3), pp. 575-580, October, 2019. © www.premierpublishers.org, ISSN: 2167-0449

Research Article

Study of Chlorophyll Mutations and Chlorophyll Content

in Young Oil Palm (Elaeis guineensis Jacq) after Gamma
*Dickson Osei Darkwah1, Essie T. Blay2, Harry M. Amoatey3, Daniel Agyei-Dwarko4, Enoch
Sapey5, Samuel Adu Osei6
Palm Research Institute, P.O. Box 74, Kade-Ghana
2 Crop
Science Department, University of Ghana, P.O. Box LG 25, Legon, Accra
3Ghana Atomic Energy Commission, P.O. Box LG 80, Legon, Accra

Chlorophyll mutations provide one of the most dependable indices for evaluating genetic effects
of mutagenic treatments and have been reported in various pulse crops by several workers. The
present investigation was undertaken to study the effects of gamma irradiation in inducing
chlorophyll mutations and as well investigate chlorophyll content after gamma radiation at 10 Gy
in M1, M2, M2M1 populations of oil palm seedlings. Chlorophyll mutations were observed 2 MAP
and chlorophyll content was determined using Chlorophyll content meter (SPAD 502) at 4, 6, 8,
10, 12 months after planting (MAP). Results revealed six chlorophyll mutants namely xantha,
chlorina, chimerical leaves, maculata, striata, and viridis. Significant differences were observed
for chlorophyll content at 4, 6, 8 MAP. Results showed that at 10 Gy of gamma irradiation,
chlorophyll mutations could be induced and chlorophyll content reduces at the initial stages of
plant growth after gamma irradiation (4-8 MAP) and later becomes normalized (10-12 MAP).

Keywords: Oil Palm, gamma rays, chlorophyll mutations, SPAD meter, chlorophyll contents


Mutation breeding in crop species can significantly Chlorophyll development seems to be controlled by many
increase genetic variability and accelerate many breeding genes located on several chromosomes (Swalingthan
programmes (Roychowdhury and Tah, 2013). 1964). Application of gamma rays may therefore cause
Mutagenesis could be used as a way of speeding up oil nuclear gene mutations or extra chromosomal deficient
palm breeding programme and creating genetic variation mutations which might result in chlorophyll deficient
due to long breeding cycle and narrow genetic base. mutations. The most dependable indices for evaluating the
Physical and chemical mutagens have been used as the efficiency of different mutagen in inducing genetic
agents of mutagenesis with onward selection of heritable variability are chlorophyll mutations.
changes of useful traits or specific genotypes for the
improvement of crop plants (Devmani et al., 2016). Chlorophyll mutations are therefore used as genetic
Induced mutations are used in plant breeding to generate markers in basic and applied research to unlock the effect
genetic variation for the development of new varieties or of different doses of mutagen in the treated plants for
novel traits. Gamma rays have been found to induce viable mutations (Patil and Rane, 2015). Chlorophyll
different types of variations than any other radiation (Patil mutations are observed easily in the M2 population.
and Rane, 2015). Gamma rays have been found to be
effective in inducing chlorophyll mutations. Chlorophyll *Corresponding Author: Dickson Osei Darkwah, CSIR-
mutations and morphological changes are some of the Oil Palm Research Institute of Ghana, P.O. Box 74,
variations observed after mutagenic treatments. Kade-Ghana Email: oseidarkwah@yahoo.com

Study of Chlorophyll Mutations and Chlorophyll Content in Young Oil Palm (Elaeis guineensis Jacq) after Gamma Irradiation
Darkwah et al. 576

According to Shu and Nakagawa (2012) mutant screening as well as radiated seeds (M1) of the same variety were
is done at the M2 stage with subsequent confirmation at obtained from the Plant Breeding Division of OPRI. Part of
the M3 stage. However Sadegh (2014) reported that the non- irradiated seeds was used as a control (M0).
though identification of mutant plants occurs at the M2
generation, there is a cogent correlation between M1 and Irradiation of seeds was done at the Radiation Technology
M2 mutation frequency caused by ionizing radiations. Centre of the Ghana Atomic Energy Commission at a
dosage of 10 Gy. Seeds were germinated using the dry
The study therefore investigates the use of gamma heat method (Hartley, 1988) for 3-4 months.
irradiation in inducing chlorophyll mutations and its effect
on leaf chlorophyll content in M1, M2, M2M1, populations. Field experiment

All the trials were conducted at OPRI-Kusi (0.6.00 N,

MATERIALS AND METHODS 001.45 W) nursery. Germinated seeds of each treatment
were sown singly in black polybags (35.6cm x 45.7cm)
Plant materials filled with a mixture of topsoil and sand in the ratio 2:1
(OPRI Recommendation) on September 2015.
Seeds were obtained from Oil Palm Research Institute
following a study carried out by Wonky- Appiah in 1976. Chlorophyll mutation
M1 palms radiated at 10 Gy showing superiority in yield
were selected. 2100 M2 seeds produced under controlled This was determined by making observations 2 months
pollination of the selected M1 were radiated again to after planting (MAP) on both irradiated and non- irradiated
produce recurrent irradiation population (M2M1) at 10 Gy. seedlings and following the description made by some
Non- irradiated seeds from a commercial oil palm variety authors below.

Table 1. Type and Descriptions of Chlorophyll Mutations

Chlorophyll Mutation Description References
1.Viridis The leaves of the seedlings are light yellowish green in colour. Gaibriyal et al. (2009)
2.Xantha The seedlings are completely yellow Patil and Rane (2015)
3.Striata The seedlings show longitudinal strips of different colours. Patil and Rane (2015)
4.Albina The leaves are white in colour Patil and Rane (2015)
5. Dark Xantha Dark yellow Gaibriyal et al. (2009)
6. Maculata Seedling showed yellow or whitish dots on leaves Kozgar (2014)
7. Tigrina Leaves yellow with green patches Kozgar (2014)
8. Chlorina Light green colour of leaves Kozgar (2014),
Devmani et al. (2016)

Determination of chlorophyll content RESULTS

Intact leaf samples from frond number 3 were used for this Chlorophyll mutations
determination. The leaf blade was surface cleaned with
distilled water and wiped. The leaf was placed between the Six different types of chlorophyll mutants were recorded for
arm and the sensor and 3 random leaf spots around the the irradiated treatments. These included xantha, chlorina,
mid-point of each leaf blade were measured for chlorophyll chimerical leaves, maculata, striata, and viridis (Plate 1).
content (Sim et al., 2015). The chlorophyll content reading For some of the treatments the leaves showed a
(ug/g FW) displayed on the screen was recorded. RCBD combination of chlorophyll mutations such as chlorina +
involving three replications was used for the experiment. maculata and viridis + chimerical leaves as indicated by
the arrows.
Ten palms were randomly selected for each of the
treatment. Data was taken at 4, 6, 8, 10, 12 MAP. Data Leaf chlorophyll content
was analysed using GENSTAT v 12
Means of each treatment were compared with those of There were significant differences P≤ 0.05 among the
controls, using Fishers least significant difference (LSD) treatments for chlorophyll content (Table 2). M1 had the
procedure. lowest leaf chlorophyll content at 4 and 6 MAP with 57%
to 60.98% reduction in chlorophyll content for these two
months. M0 had the highest chlorophyll content at 4, 6, 8
and 10 MAP. At 10 MAP, there were no significant
differences P≤ 0.05 for leaf chlorophyll content for all the

Study of Chlorophyll Mutations and Chlorophyll Content in Young Oil Palm (Elaeis guineensis Jacq) after Gamma Irradiation
Int. J. Plant Breed. Crop Sci. 577

Chlorophyll mutant: Chimera Chlorophyll mutant: Chimera

Population : M2 Population : M2

Chlorophyll mutant: Striata Chlorophyll mutant: Viridis + Chimera

Population: M1 Population: M1

Chlorophyll mutant: Xantha Chlorophyll mutant: Chlorina

Population: M1 Population: M1

Chlorophyll mutant: Maculata Chlorophyll mutant: Maculata+ Chlorina

Population: M2M1 Population: M2M1

Study of Chlorophyll Mutations and Chlorophyll Content in Young Oil Palm (Elaeis guineensis Jacq) after Gamma Irradiation
Darkwah et al. 578

Non - irradiated seedlings

Plate 1. Chlorophyll mutations following gamma irradiation in oil palm seedlings

Table 2. Effects of Gamma Irradiation on Chlorophyll content (ug/g FW)

Treatments Months After Planting
4 6 8 10 12
Mo 49.74a 51.8a 48.89a 48.88a 49.64a
M1 28.15 c 31.59 c 44.68 a 42.44 a 50.68a
M2 38.07 b 44.48 b 39.22 ab 43.48 a 48.82a
M2M1 36.52 b 48.27 a 38.17 ab 45.42 a 46.13a
Means with the same letter (s) are indicative of significant difference at 5% Probability level.

DISCUSSIONS Viridis chlorophyll mutations were more than the others.

This has been attributed to the participation of polygenes
Chlorophyll mutations in chlorophyll formation (Akpte et al., 2006; Ambarka et al.,
Chlorophyll mutations are reliable indicators for estimating
the efficiency and effectiveness of various mutagens; be it Leaf chlorophyll content
physical or chemical in inducing genetic variability for crop
improvement purposes (Devmani et al., 2016; Gaibriyal et Differences in plant chlorophyll content were highly
al., 2009). For basic and applied research purposes, significant among the treatments. M0 had the highest value
chlorophyll mutations are used as genetic markers followed by M2 and M2M1 with the least being M1 at 4MAP.
(Devmani et al., 2016). They also reported that low or Bornman (1989), reported that the chloroplast is the
moderate doses of gamma irradiation give a higher significant point of destruction following gamma irradiation,
frequency of chlorophyll mutations and that increasing the which eventually leads to loss of chloroplast integrity
dosage does not increase the mutagenic efficiency. affecting photosynthetic ability. The reduction of the
Observation of chlorophyll mutants is expected to occur in chlorophyll content is as a result of selective destruction of
M2 generation (Shu and Nakagawa (2012). However chlorophyll ‘b’ biosynthesis or degradation of chlorophyll ‘b’
Sadegh (2014) reported that though identification of precursors (Borzouei et al., 2013). Sreedhar et al. (2013)
mutant plants occurs at the M2 generation, there is a from transmission electron microscopy concluded that the
cogent correlation between M1 and M2 mutation frequency organized pattern of grana and stroma thylakoids was lost
caused by ionizing radiations. In this study chlorophyll and thylakoids appeared slightly widened after gamma
mutations were identified at seedling stage in all the three irradiation. Wu et al.(2007) noticed that lower chlorophyll
treatments namely M1, M2, and M2M1. Six chlorophyll concentration at the early stages of rice might be due to
mutants were identified namely xantha, chlorina, parallel degradation of pigments and pigment binding
maculata, viridis, striata and chimerical leaves. The viridis proteins of the photosynthetic apparatus as well as
was more frequent than all the other chlorophyll mutations. delayed chloroplast development. Delayed chloroplast
However chlorophyll mutations were not observed in the development might lead to a slow accumulation of
control seedlings. Chlorophyll mutations observed in this chlorophyll in irradiated oil palm. Kiong et al. (2008) also
studies are in agreement with other authors; Gaibriyal et confirmed a derangement in the motif of the grana and
al.(2009) and Usharani and Kumar (2013) in blackgram, thylakoid and destruction of the chloroplast following
Devmani et al. (2016) in cowpea, Usharani and Kumar gamma irradiation which causes injury to the chlorophyll
(2015) in urdbean, Kozgar (2014) in chickpea and Solanki pigments in plants.
(2005) in lentil.

Study of Chlorophyll Mutations and Chlorophyll Content in Young Oil Palm (Elaeis guineensis Jacq) after Gamma Irradiation
Int. J. Plant Breed. Crop Sci. 579

The results obtained in this study are in harmony with germination and seedling growth. Emirates Journal of
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Copyright: © 2019 Darkwah et al. This is an open-access
Impaired Chlorophyllide Esterification in Chlorophyll
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Biosynthesis. Plant Physiology, 145(1) : 29–40.
Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium,
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Study of Chlorophyll Mutations and Chlorophyll Content in Young Oil Palm (Elaeis guineensis Jacq) after Gamma Irradiation