RESEARCH PROPOSAL

For Ph.D. degree program

Title: ALLEVIATION OF ADVERSE EFFECT OF DROUGHT STRESS IN MAIZE BY

NATURAL AND SYNTHETIC GROWTH PROMOTERS

PERSONAL DETAILS

Full name: Zaid

Father’s name: Abdul Hameed
Nationality: Pakistani
Permanent mailing address: Hafiz
Cell#: +92
Email addresses: zs.chachar@gmail.com
Background:

Maize is an important cereal crop grown all over the world. It is the third most important cereal
after wheat and rice all over the world serving as staple food for many countries (Frova et al., 1999).
In Pakistan, maize is grown on an area of 950,000 ha with the production of 3,487,000 tones and
average grain yield of 3671 kg ha-1 (Government of Pakistan, 2010).

In Pakistan diverse maize genotypes i.e., single cross and double-cross hybrids, synthetics and
composites are being planted. The responses of all these genotypes vary to different
agromanagement practices, particularly water and nutrient. It is estimated that over the next 35
years the demand for maize in the developing world will double. Maize being the highest yielding
cereal crop in the world is a significant importance for countries like Pakistan, where rapidly
increasing population has already out stripped the available food supplies.

Water stress induce numerous physiological and biochemical changes in plants like reduced leaf
size, stem extension, root proliferation, reduced water use efficiency), negatively affect the regular
plant metabolic activities (Lawlor & Cornic, 2002), inhibition of enzymatic activities (Ashraf et
al., 1995), ionic imbalance and disturbances in solute accumulation (Khan et al., 1999) or a
combination of all these factors. In maize, drought reduces leaf area, leaf chlorophyll contents,
photosynthesis and ultimately lowers the grain yield (Athar & Ashraf, 2005). At flowering, drought
widens the anthesis-silking interval (ASI) in maize, which severely reduces the kernel set
(Emeadeas et al., 2000). Under drought leaf senescence is also accelerated to decrease the canopy
size (Moony & Duplesis, 1970) severely affecting the crop yield. However delayed leaf senescence
affects positively for reducing the harmful effects of drought on crop yield (Rivero et al., 2007).

Different approaches are applied to mitigate the drought effects, which include the evaluation of
stress tolerant plants by selection of stress-resistant varieties (Ahloowalia et al., 2004), in vitro
selection, use of plant growth hormones (ABA, GA, cytokinin, SA), antioxidants (ascorbic acid,
H2O2) and osmoprotectants such as proline, glycine betaine as foliar application and seed treatment
(Senaratna et al., 2000; Farooq et al., 2009). Among growth regulators, Cytokinins play a critical
role in promotion of cell division, chlorophyll biosynthesis and modification in apical dominance
in plants (Taiz & Zeiger, 2006). Cytokinin application under abiotic stressful conditions can delay
the leaf senescence directly by scavenging reactive oxygen species (Miller, 1992; Grossman &
Leshem, 2006). When maize plants genetically modified with IPT gene subjected to drought stress,
then IPT gene was overexpressed and induced the endogenous cytokinin production, which resulted
in stay green characteristic and ameliorated the adverse effects of drought (Robson et al., 2004).
Cytokinins are commercially available in the form of benzyl amino purine (BAP) or Trans-Zeatin,
but their use is very much costly. Moringa (Moringa oleifera L.) is a tree commonly known as “the
miracle tree”. It is well known for its nutritional and medicinal properties. Leaves of moringa are
rich in vitamins (A, B, C), essential minerals (K, Ca, Fe), antioxidants (Ascorbate, Phenolics),
proteins and zeatin (Foidle et al., 2001). Hence, its leaf extract either prepared in 80% ethanol or
in water contains growth-enhancing substances and can be used as a natural source of growth
promoter (Fuglie, 2000), which may reduce the adverse effects of drought stress on maize by
delaying the leaf senescence and scavenging the reactive oxygen species. Plants treated with this
spray exhibited more pest and disease resistance, vigorous life-span, heavier roots, stems and
leaves, bigger fruits, higher sugar levels and an overall 20-35% increase in yield (Makkar & Becker,
1996).

Few reportings are available on the use of alternate naturally occurring leaf extracts for the
mitigation of stressful conditions especially drought. Use of moringa as a growth enhancer was
found a potential source to mitigate the deleterious effects of drought. Therefore, the present study
will be organized to evaluate the potential of moringa leaf extract as growth enhancer and delaying
the leaf senescence of maize under drought stress conditions, while exploring improved drought
tolerance on the physiological and biochemical basis.

Objectives of the study

 The optimum concentration of moringa leaf extract will be optimized to enhance drought
tolerance of maize.
 Effect of moringa leaf extract on Physiological, biochemical and morphological changes in
maize will be studied while subjected to different water stress level.
 Potential of moringa leaf extract might have some significant effect on plant growth
enhancement and delaying the leaf senescence of maize under drought stress conditions.

Material and methodology

This study will be conducted under net house. Maize seed will be collected from relevant seed
department and will be used as planting material. The experiment will be laid out in Completely
Randomized Design (CRD) with factorial arrangements using three replications. Maize seedlings
will grown in the clay pots containing soil and compost (1:1). Four seeds will be sown in one pot.
At four leaf stage thinning will done and one plant per pot was maintained till physiological
maturity. Drought stress will be imposed after four weeks of sowing when seedlings attained the
four leaf stage. Three drought levels i.e., 100% field capacity (FC), 75% FC and 50% FC will
maintained on gravimetric basis
Three foliar treatments viz Water spray (Control), MLE (1:30) Optimized dilution level by
Iftikhar (2009) and Benzyl amino purine (BAP) @ 50 mg L-1; application will be started one week
after imposition of drought stress, onward it will be sprayed on weekly basis. For each spray 25
mL BAP or MLE will be applied per plant. Moringa leaf extract was prepared by grinding the
young frozen leaves of moringa. The extract prepared will be further diluted 30 times by adding
the distilled water. For the preparation of BAP solution, BAP will first dissolved in ethanol and
then distilled water will be added to get the 50 mg L-1 concentration.

Major studies will be focus on

Cell membrane thermos-stability, Plant water relations, Biochemical analysis (Chlorophyll a and b
and total chlorophyll, carotenoid), Antioxidant activities of key antioxidant such as SOD, POD,
CAT, APX and GR with be studied, moreover, phenolic compounds, osmoprotectant (Proline and
Glycine betaine) Hormones (ABA, GA, Cytokine, IAA and jasmonate. Furthermore plant growth
and yield related agronomical studies will also be conducted during growth and at harvesting stage.
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