Iqbal Hussain | Pakistan Agricultural Research Council (original) (raw)

Papers by Iqbal Hussain

Zinnias originates from Mexico and has high demand in the floriculture market. The need to reduce... more Zinnias originates from Mexico and has high demand in the floriculture market. The need to reduce reliance on chemical fertilizers has highlighted the importance of sustainable alternatives like organic manures. Compost application in horticulture is gaining attention as a sustainable alternative to chemical fertilizers, aiming to enhance soil health and plant growth. However, there is a limited understanding of its effects on the growth. This study investigates the effects of compost on the growth, flowering attributes, and vase life of Zinnia elegans to improve cultivation techniques and promote sustainable horticultural practices. The experiment used an RCBD with two factors. When they reached 3-5 true leaves, plantlets were transplanted into 12-inch pots with compost varying concentrations, including Control, 25%, 50%, and 100% with garden soil. Changing compost concentrations and zinnia varieties affected numerous factors strongly. Plant height, for instance, varied significantly, with Benary's Giant cultivar reaching the tallest (72.04 cm) under 50% compost, mirroring the stem diameter's peak at 1.01 cm with the same compost level. Compost levels and zinnia varieties also notably impacted leaf count, lateral branches, leaf area, flower count, and flower attributes like diameter and stalk length. The study also found that 50% of compost extended petal senescence survival. Compost and zinnia interaction affected the flower's fresh weight and vase life. The interaction cultivar Benary,s Giant with 50% compost in garden soil was optimal for zinnia growth and flowering. This study will provide the basics for floriculture.

EFFECTS OF GROWTH REGULATOR ON INVITRO PROPAGATION OF LILIUM USING BULB SCALE

Lilium spp. is the most significant decorative plant and has high demand in the floriculture mark... more Lilium spp. is the most significant decorative plant and has high demand in the floriculture market. We studied the impact of different growth regulators on culture initiation, shoot proliferation, and root formation of Lilium for disease-free plants from in vitro propagation. The explant basal scale was used for culture initiation, and varying concentrations of BAP (6-Benzylaminopurine) alone and in combination with IAA (Indole-3-acetic acid) showed different responses. It was observed that MS media supplemented with BAP at 1.5 mg/l and IAA at 1.0 mg/l exhibited the maximum number of shoots, i.e., 4.3 shoots and 7 cm length of shoots, which were attained in four weeks. Several concentrations of BAP and Kinetin (KIN) supplemented with MS medium alone or in combination with BAP with IAA, IBA, and NAA were used for shoot proliferation. The results revealed that all treatments resulted in 100% shoot development, with significant variability in the number of shoots and shoot length. MS supplemented with 1.5 mg BAP and 1.0 mg KIN showed the maximum 7.4 number of shoots, with an average shoot length of 7.7 cm. The maximum 11 roots observed in MS supplemented with 1.5mg IBA and the longest roots 4.4cm found in MS with 2mg IAA, and the rooted plantlets were hardened in peat moss media, had the highest ex-vitro survival rate. These results highlight the significance of optimized combinations of growth regulators for improving micropropagation efficiency in the horticulture industry.

Evaluation of the Transgenic Potato Plantlets in Hydroponic Culture for Salt Tolerance

The agricultural crop yield has been lessening due to the adverse effect of global warming and cl... more The agricultural crop yield has been lessening due to the adverse effect of global warming and climate change. Salts stress is major yield devastating factor in crops. Hydroponics is an approach of growing plants in aqueous solution carrying basic nutrients without soil. In this experiment evaluation of plantlets of transgenic potato varieties Asterix and Sante having AtNHX1 gene along with their wild types (non-transgenic) was carried out at National Institute for Genomics and Advanced Biotechnology (NIGAB), National Agricultural Research Centre (NARC), Islamabad in the screen house conditions. The transgenic and control plants were tested at different NaCl levels 0.0, 20, 40 and 60 mM NaCl under hydroponic conditions. Both the transgenic potato lines not only survived successfully but also showed greater tolerance to salt stress even at 60 mM NaCl for plant height, number of nodes per plant, root length, root weight, shoot weight and total chlorophyll contents. On the other hand, the non-transformed plantlets (control) of Asterix and Sante could not survive even at 20 mM NaCl stress.

In vitro regeneration in maize, 2017

In vitro regeneration system of six maize inbred lines (EH1, EH2, EH3, EH6, EH8, EH9) for direct ... more In vitro regeneration system of six maize inbred lines (EH1, EH2, EH3, EH6, EH8, EH9) for direct shoot multiplication was established using 1st collar node as explants of one-week-old in-vitro plantlets. The Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP) in combination with Indole acetic acid (IAA) and GA3 were used. Results showed that the addition of BAP, IAA, and GA3 greatly enhanced the formation of multiple shoots in the tested lines. Significant differences were observed between the concentration of growth hormone and inbred lines. Among the six treated inbred lines, EH6 performed best for direct multiple shoot regeneration. Maximum number of shoots (11 shoots per explant) was observed on the medium containing BAP (0.7 mg/l) in combination with IAA (0.2mg/l) and GA3 (0.5 mg/l). Regenerated shoots were rooted in MS supplemented with 2 mg/l NAA. The rooted in vitro raised plantlets were acclimatized and successfully transferred to the greenhouse. The regeneration protocol developed in this study can play an important role in the mass propagation of elite genotypes of maize after successful transformation of the desired gene.

Effect Of Different Treatmentsonthe Safe Removalof Seed Dormancyin, 2014

Abstract: Sunflower seed dormancy is a major drawback in terms of proficient seed production. The... more Abstract: Sunflower seed dormancy is a major drawback in terms of proficient seed production. The
high level of variation in the seed dormancy of sunflower was observed in the previous studies. Eight
different treatments were used to determine seed dormancy of sunflower hybrid Hysun-33. The effect
of light and dark was also observed on the germination of treated and control seeds. Significant
differences were observed among the different treatments. Light and dark effect has also significant on
seed germination. The treated seeds placed under light gave more mean value of seed germination as
compared to dark condition. Results showed that seeds treated with 1g Thiourea for 15 min and soaked
in water for overnight, placed under light condition gave maximum seed germination (90.33) after ten
days followed by seeds treated with 10% of GA3 for 15 min and soaked in water for overnight gave
mean value of seed germination (80.00). The effect of dark was also observed in the seeds that were
treated with hot water (80 oC for 1 min) and gave maximum seed germination of (42.33) after ten days
of treatment. No any seed germination was seen in control. This protocol of breaking seed dormancy of
sunflower will be helpful for the researchers on different aspects.
Key Words: Seed Dormancy, Sunflower, Thiourea, GA3, Light, Dark condition.

Inter Simple Sequence Repeat-Based Genetic Divergence and Varietal Identification of Banana in Pakistan, 2022

This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY

Simple Sequence Repeats-Based Genetic Characterization and Varietal Identification of Potato Varieties Grown in Pakistan, 2022

AN EFFICIENT AND REPRODUCIBLE TISSUE CULTURE PROCEDURE FOR CALLUS INDUCTION AND MULTIPLE SHOOTS REGENERATION IN GROUNDNUT (Arachis hypogaea L, 2020

This study was carried out to optimize in vitro regeneration system of groundnut through tissue c... more This study was carried out to optimize in vitro regeneration system of groundnut through tissue culture technology that can provide good environment in terms of food security. The two varieties of groundnut (Golden and BARI 2001) were used and an effective in vitro regeneration system was established for selecting the best responsive variety to tissue culture using embryo as an explant sources. MS media having 5.5 mg/l BAP and 1.5 mg/l NAA was found to be best by producing the highest callus induction frequencies (86 & 78%) in Golden and BARI 2001, respectively. Similarly, the maximum multiple shoots/ plant (9 & 6) with optimum length of shoots (4.8 & 4.1 cm) was obtained with the application of 4 mg/l BAP along with 1 mg/l NAA and 1.1 mg/l TDZ in Golden and BARI 2001, respectively. In case of roots initiation, the highest root frequency (90 & 76%) having optimum roots/shoots (8.3 & 6.2) and substantial root length (8.1 & 7.0 cm) was obtained in Golden followed by BARI 2001 variety on MS media having 1 mg/l phytohormone IBA. Results showed that callus and multiple shoot induction and in vitro rooting, varied with different hormonal combinations in the media and the genotype that might be due to genetic variations between the varieties. This study delivers a base line for efficient in vitro propagation and genetic transformation in elite groundnut genotypes for desirable characteristics.

PJAS, 2019

Potato is an important food crop of the world. Different viral, bacterial and fungal pathogens ca... more Potato is an important food crop of the world. Different viral, bacterial and fungal pathogens cause heavy economic losses of
this crop every year. Potato has complex genetic makeup due to which induction of disease resistance through conventional
breeding is difficult. Genetic manipulation through different transformation techniques is more precise and successful tool. In
the present study different factors were investigated, which have an influence on potato transformation. The optimal dose of
cefotaxime was found 500 mg/l which did not affected the growth of the potato tissues. The explants treated with
Agrobacterium in the presence of acetosyringone resulted in higher frequency of transformation as compared to the explant
without it. Two minutes time for co infection was found appropriate for optimum transformation efficiency. The two days cocultivation period along with 7 days preselection was found suitable for potato transformation. The putative tranformants
regenerated on MS medium supplemented with 20 mg/L hygromycine and 500 mg/L cefotaxime, from nodal explants while
the non-transformed tissue turned brown and gradually died after two or three sub culturing on the selection media containing
selective antibiotic hygromycin. The shoots obtained on selection media shifted on root induction media supplemented with
similar concentrations of hygromycin and cefotaxime, resulted complete plantlet formation after 10 days. It was observed that
all the hygromycin positive plants also exhibited positive bands of desired size of 823 bp for chitinase gene, suggesting cotransformation of both genes in transformed plants.
Keywords: Solanum tuberosum, chitin, hydrolytic enzyme, fungal pathogens.

MOLECULAR CHARACTERIZATION OF Fusarium oxysporum F. SP. CUBENSE (FOC) TROPICAL RACE 4 CAUSING PANAMA DISEASE IN CAVENDISH BANANA IN PAKISTAN, 2017

Fusarium wilt or Panama disease poses a great risk to banana production worldwide and is caused b... more Fusarium wilt or Panama disease poses a great risk to banana production worldwide and is caused by Fusarium oxysporum f.
sp. Cubense (Foc). Its tropical race 4 (TR4) is a major problem for Cavendish banana growing areas. Basrai (Dwarf
Cavendish) is the major cultivated variety in Pakistan and now recently been reported to be affected by Foc TR4 in certain
areas in the Sindh province. To genetically characterize, three independent isolates (P.TH1, P.TH2 and P.Th3) were sampled
from different fields in district Thatta and fungus was isolated on potato dextrose agar. The pure fungal isolates were used to
establish Koch’s postulates by testing their pathogenicity on tissue cultured Cavendish banana. A diagnostic PCR assay was
optimized and used to sequence a reported molecular marker for specific detection of Foc TR4 and to sequence 28S-18S
ribosomal RNA intergenic spacer region (IGS) from these local isolates. The phylogenetic of Pakistani isolates suggests that
they belong to previously characterized lineage V of tropical race 4 of F. oxysporum cubense with 99% bootstrap support
and show close relationship with Indonesian and Malaysian Foc TR4 isolates. The rapid spread of this disease in different
banana growing areas of the Sindh is alarming and necessitates effective control of this through chemical and biocontrol, if
remains unchecked, Foc TR4 can raise havoc to banana industry in Pakistan and would be devastating economically.
Keywords: Panama disease, Fusarium oxysporum f. sp. Cubense, tropical race 4, banana wilt, phylogenetic analysis

This study was aimed to develop a simple, comprehensive and efficiently repetitive protocol for m... more This study was aimed to develop a simple, comprehensive and efficiently repetitive protocol for micropropagation of banana (Musa sapientum L.) using shoot meristem. After growing in-vitro cultures on different hormonal combinations, Murashige and Skoog's medium supplemented with BAP and IAA (5.0 + 1.0 mg/l, respectively) and 10% CW were found to be most efficient and productive combination for shoot proliferation. Antibiotic (cefotaxime) was used to check the endogenously born bacterial contamination. Initiation of shoot bud and establishment of culture from shoot meristem was achieved on liquid media. Further shoot proliferation of cultures up to 3 batches of 15 days each was achieved on the agarose gel solidified hormonal supplemented media after culture establishment. The proliferated shoots were excised and transferred to different root induction media, which resultantly showed that MS media supplemented with IAA (2mg/l), was the most efficient root inducing media. Rooted plantlets after primary and secondary hardening were transferred to the green house. Finally, these disease free plants were successfully established in soil.

The present study was planned to evaluate the transgenic potato plants containing chitinase (RCG-... more The present study was planned to evaluate the transgenic potato plants containing chitinase (RCG-3) gene transformed through Agro bacterium for resistance against Rhizoctonia Solani and Fusarium solani. In vitro, plants were cultured for multiplication and then pathogenicity of transgenic potato plants harboring chitinase gene were carried out with fungal pathogens (R. solani and F. solani). The fungal pathogens were isolated from infected tubers and purified by single spore technique. For multiplication of pathogens wheat straw method was used for R.solani and sorghum seed method was used for F.solani. The inoculation of fungal infection (R. solani) and (F. solani) transgenic and non-transgenic plants were performed in laboratory and controlled greenhouse conditions (Containment). Results showed that transgenic potato plants containing chitinase (RCG-3) gene transformed through Agrobacterium showed resistance against R. Solani and F. solani compared to non-transgenic plants. It was concluded that rice chitinase gene expression in potato confers enhanced resistance against two major fungal diseases of potato in Pakistan.

In vitro response and its relationship with different varieties, explants and media were investig... more In vitro response and its relationship with different varieties, explants and media were investigated in potato (Solanum tuberosum). Direct In vitro regeneration protocol from diverse explant source is a prerequisite for transformation studies. Three potato cultivars viz., Cardinal, Altamash and Diamont were selected for in vitro responses. High regeneration and morphogenic potential of different explants i.e., shoot tips, leaf discs, nodes and internodes have been tested for direct regeneration. Basal media was Murashige & Skoog and different hormonal combinations of benzyl adenine and indoleacetic acid were supplemented. Statistical analysis showed that explant source had significant effect on direct regeneration and the nodal explants had maximum regeneration. The number of shoots obtained from node was 17.6 from Cardinal followed by Diamont 14.3 and Altamash 9.0. Shoot apices also resulted in shoot regeneration comparatively better than leaf discs and internodal explants but lesser than from nodes. Most suitable medium was MS with 2.0 mg/l BAP and IAA @ 0.5 mg/l giving maximum regeneration. It was also observed that interaction of cultivars with explant and media is highly significant at P 1.0%.

Protocols for in vitro tuberization of potato (Solanum tuberosum L.) variety Cardinal were studie... more Protocols for in vitro tuberization of potato (Solanum tuberosum L.) variety Cardinal were studied. Virus free cultures were multiplied on Murashige & Skoog (1962) medium containing 1.0 mg/l GA 3. When the number of shoots was maximum the cultures were shifted on tuber induction medium. In vitro tuberization media consisted of MS along with different concentrations of CCC, BAP and Sucrose. It was observed that 200 mg/l of CCC or 90g/l sucrose in the mediums resulted in maximum tuber induction (16.5 and 15.6 tubers/flask respectively) followed by BAP @ 4.0mg/l producing 9.0 tubers/flask. It was also observed that complete darkness was essential for tuber induction.

HortScience, 2007

The effect of benzylaminopurine (BAP) and kinetin alone and in combination with indole 3-acetic a... more The effect of benzylaminopurine (BAP) and kinetin alone and in combination with indole 3-acetic acid (IAA) on shoot proliferation of ‘Basrai’ (Musa spp., AAA group) was investigated. Shoot tips (4–6 mm) were excised from field-grown suckers to initiate the cultures. Concentrations of BAP and kinetin ranged from 0.0 to 8.0 mg·L−1 each on solid or in liquid MS medium. When liquid medium was used, cultures were agitated continuously on an orbital shaker moving at 40 rpm. Three subculture regimes were employed; after each subculture, the number of shoots regenerated from each explant was counted. The results showed that the multiplication rate was significantly (P ≤ 0.05) dependent upon cytokinin type, its concentration, and type of medium used. The maximum number of shoots regenerated from a single shoot tip was achieved in liquid MS medium containing 4.0 mg·L−1 BAP. There was no significant difference between liquid and solid medium when kinetin was used; however, kinetin at 4.0 mg·L−...

AFRICAN JOURNAL OF BIOTECHNOLOGY, 2012

A voluminous body of research has reported the establishment of efficient protocols for sugarcane... more A voluminous body of research has reported the establishment of efficient protocols for sugarcane multiplication through tissue culture. However, a reliable and reproducible in vitro plant production system remains obscured. Furthermore, validation of virus free nature of in vitro plants using molecular techniques is the most challenging one. Considering the need for high yielding cultivars due to land and constraints, this study was devised for mass multiplication of high yielding elite cultivars of sugarcane viz. HSF-240, YT-55 and YT-53. Use of 100% Clorox for surface sterilization of apical and lateral buds, and of cefotaxime (500 mg L-1) for controlling bacterial contaminants revealed complete sterilization of field grown explants. Culture initiation was dependent upon plant growth regulators (PGRs), genotype and type of explants. The highest shoot initiation frequency of 96% was obtained with combination of four plant growth regulators (0.1 mg L-1 BAP), (0.1 mg L-1 NAA), (0.1 mg L-1 Kn) and (0.1 mg L-1 GA 3). Maximum shoot number (17.4) was exhibited by HSF-240 on MS media when the concentrations of BAP, Kn and GA 3 were increased to 1 mg L-1 in combination with NAA (0.25 mg L-1) indicating preference for higher concentrations of PGRs. Half-strength MS media with 6% sucrose resulted in increased root length (9.2 cm) and root number (20.5) Hardening efficiency of 98.6% was achievable in sandy clay loam soil. Two steps reverse transcription PCR (RT-PCR) was successfully employed for detection of sugarcane mosaic virus (SCMV) in in vitro plants. These results have implications for understanding optimum conditions for in vitro mass production of sugarcane plants, molecular detection of SCMV in in vitro raised plants, and stable genetic transformation studies.