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THIS STudy was aimed to induce genetic variation toward drought tolerance after treatment with EMS as a mutagen to two maize inbred lines and their single hybrid. The experiment was carried out at Abo-Ghraib Research Station, Baghdad during the spring season of 2019. The experiment included three irrigation intervals 7, 9 and 11, four concentrations of EMS (0, 50, 60, and 70 mM) and three maize genotypes distributed in three replications using randomized complete block design (RCBD) with split-split plot arrangement which included three irrigation intervals 7, 9 and 11 days occupied the main plot, while the sub plots included three maize genotypes, and four EMS concentrations which were distributed in sub-sub plots. Results were revealed significant differences among EMS treatments and irrigation interval treatments. The inbred line 5 in control treatment showed a significant increase in days for both anthesis and silking (70.11 and 71.94) days, respectively. Plants of Nahrain hybrid also showed a super performance in plant height, leaf area, and number of rows ear-1, with a mean of 154.47cm, 746.31cm2 and 17.83 row ear-1, respectively. The irrigation intervals of 9 and 11 days were significantly decreased the mean performance of genotypes, compared to the shortest irrigation interval (7 days) which had highest mean in most traits. None treated plants (0 mM EMS) had a significant effect on the performance of most studied traits, except the anthesis and silking time, which gave the highest values under the concentration 60 mM with 69.11 and 70.48 days, respectively. Hybrid plants interacted significantly with the EMS concentration of 0 mM producing the maximal means of plant height and leaf area (169.56cm and 759.78cm2 respectively). On the other hand, the hybrid was significantly interacted with the shortest irrigation interval in the mean traits of plant's height and leaf area (163.50 cm and 757 cm2, respectively). The triple interaction between hybrids, shortest irrigation interval and EMS concentration of 60 mM had significantly increased leaf area reached 769.67cm2, meanwhile the triple interaction between hybrid plants, shortest irrigation interval, and the EMS concentration of 0 mM recorded the highest value of plant yield (199.67g). It can be concluded from the current study that the genotypes differed in their response to 9 and 11 days irrigation intervals, with a negative effect of the later interval on the vegetative and yield traits of maize compared with shorter irrigation interval.

Dehydration is one of the most important determinants to plant growth and productivity in general, and maize in particular. Therefore, the current study aimed to induce genetic variation for drought tolerance in two maize inbred lines and their F1 hybrid with aid of ethyl methanesulfonate EMS mutagenesis. Furthermore, the simultaneous effect of EMS was investigated in the expression of MnSOD and Cu/Zn-SOD genes. The study was conducted in the fields of Abu Gharib Station-Agricultural Research Department- Baghdad in the spring season of 2019. The results showed that the genotypes differed significantly in the levels of the expression of both MnSOD and Cu/Zn-SOD genes. The gene pool of 18 and 5 parental lines and their single hybrid (Nahrain) reflected on the expression level of Cu/Zn-SOD gene, that produced 39.54, 17.63 and 24.37 copies, for the three genotypes, respectively at the 7-day irrigation interval. There was an increase in the expression level of the previously mentioned gene in most genotypes at a concentration of 70 mM EMS under the longest irrigation interval (11 days). The plants of inbred 18 were the best at the level of expression of the Cu/Zn-SOD gene with 18.61 copies, followed by the single hybrid and the parental inbred 5 with 11.99 and 7.79 copies, respectively. The investigate genotypes revealed clear differences in the expression of MnSOD gene in response to the two studied factors, as hybrid plants obtained the highest level of gene expression reached 120.59 copies at the EMS concentration of 70 mM and irrigated with the longest interval. It was followed by untreated plants of inbred 18 (control treatment) with 98.33 copies, then inbred 5 which produced 61.92 copies of the same gene at the EMS concentration of 60 mM and under the shorter irrigation interval (7 days). The overall performance of the F1 hybrid and parental line 18 in respect of MnSOD gene expression was nearly identical with 41.83 and 41.81 copies, respectively.

The present study investigated the effect of gamma irradiation on biomass formation and yield of pharmacologically relevant secondary metabolites in callus cultures induced from different seedling parts (leaf, stem and root). Calli induction and maintenance were performed on MS medium supplemented with 0.5 mg L−1 IAA and 0.4 mg L−1 TDZ and all parameters were recorded after the third 21 days successive subculture post-elicitation. Fresh callus biomass (FCB) expressed as growth index (Gi) was investigated and the higher Gi value (1109%) was recorded on callus culture from leaf explants and irradiated with 10 Gy dose, as compared to untreated control (757%). H. triquetrifolium callus cultures exhibited quantitatively different phenolic compounds (p-OH-benzoic and chlorogenic acid, epicatechin) and naphtodiantrones (hypericin and pseudohypericin). The chromatographic analysis of callus extracts revealed that after gamma rays elicitation with 10 Gy dose, the higher amount of p-OH-benzoic acid was recorded on calli from leaf explant (4.35 mg 100 g−1 DW material) while root callus recorded the highest amount of chlorogenic acid (12.91 mg 100 g−1) over the control (3.55 mg 100 g−1 and 10.22 mg 100 g−1, respectively). The 10 Gy and 20 Gy irradiation doses stimulate the epicatechin accumulation on calli from leaf and stem (126.39 and 148.80 mg 100 g−1) compared to the control samples (98.81 and 101.72 respectively). In this study, small amount of hypericin and pseudohypericin were identified and quantified on irradiated callus cultures initiated from stem and leaf of Hypericum triquetrifolium, but not in root calli. Callus induced from leaf and irradiated with 10 Gy showed the higher amounts of hypericin and pseudohypericin content (0.29 and 4.00 mg 100 g−1 DW material). Phytochemical screening carried out on different elicited callus cultures variants reveal, for the first time, the stimulatory effects of gamma irradiation on the production of phenolic compounds and naphtodiantrones in Hypericum triquetrifolium Turra. The results of this study are interesting and offer an innovative approach of elicitation, not yet widely studied for the Hypericum genus.

To explore the potential for in vitro rapid regeneration of Spanish dagger (Yucca gloriosa 'Variegata'), different concentrations of 6-Benzyladenine (BA), 1-naphthaleneacetic acid (NAA) and combinations of both were evaluated for callus induction initiated on leaf and bud (terminal and axillary buds) explants using Murashige and Skoog (MS) medium. Callus response induction percentage, fresh weight, color and texture of the callus were assessed after 1.5 and 6.0 months in culture. The appropriate medium for callus initiation on leaf explants was MS medium supplemented with 6.0 mg/L NAA. A combination of 0.2 mg/L BA and 1.5 mg/L NAA also exhibited a remarkable callus induction on bud explants. Effect of thidiazuron (TDZ) addition to the culture medium supplemented with BA and NAA at various concentrations (0.0, 0.1, 0.5 or 1.0 mg/L) was investigated. Faster callus growth and more callus proliferationon from bud explants were observed in MS medium supplemented with 0.1 mg/L BA + 1.5 mg/L NAA + 1 mg/L TDZ. Results showed that TDZ was more effective on callus induction than BA or NAA, however some combinations gave direct shoot regeneration. Calli derived from bud explants were whitish to creamy and compact, while calli derived from leaf explants were greenish.

This study was aimed to micropropagate Lisianthus Eustoma grandiflora plant which is one of the important cut flowers. The study included many experiments, at the initiation stage, the effect of the two auxins Indole butyric acid (IBA) and Naphthalene acetic acid (NAA) supplemented to Murashige and Skoog medium (MS) containing 0.3 mg/l of Benzyladenine (BA) were investigated for their effect on mean number of initiated shoots and length. type of explant were used namely, leaf discs. The effect of different strengths of MS and B5 media on mean number of multiplied shoots was also studied. During acclimatization, different mixture ratios of peat moss and river sand were examined in their effect on plantlets survival. Results showed that IBA achieved the highest mean shoot number formed on the three explants types under investigation reached 6.8 shoots with a mean length of 2.75 cm. Maximum mean shoot number reached 7.3 at the concentration 0.1 mg/l of IBA while maximum mean shoot number reached 6.4 at the concentration 0.0 mg/l of NAA. Results also revealed that MS medium was better than B5 medium in sustaining shoots yielding 8.8 and 6.6 for both media respectively. A significant reduction in mean number of shoots occurred with reducing the medium salt strength. Shoots transferred to MS medium supplemented with 1.0 mg/l of IBA produced the highest mean root number, length and rooting percentage reached 5.5 roots, 2.5 cm and 70% respectively while rooting percentage did not exceed 20% when the same medium was supplemented with 1.0 mg/l of NAA. Plantlets were successfully acclimatized achiving 90% survival when grown on agricultural medium consisted of river sand : peat moss (2:1)v/v.

This project aimed to increase the production of some secondary metabolites using chemical elicitors in tissue cultures of Solanum nigrum L. plants. The quality and quantity of phytochemicals were estimated using methanolic extracts of dried leaves and callus were analyzed using HPLC. Callus was initiated from leaf discs cultured on Murashig and Skoog (MS) medium supplemented with Naphthalene acetic acid (NAA) at concentrations of 1.0, 2.0, 3.0, 4.0 or 5.0 mg/l and BA at 0.5, 1.0, 1.5, 2.0 or 2.5 mg/l for S. nigrum callus initiation. The same combination was used for callus maintenance for the plant. The results showed an increase in the concentration of secondary metabolites in methanol extracts induced from leaves. Callus cultures induced from leaf discs were treated with some chemical stimuli such as jasmonic acid and salicylic acid, (2,4,6, 8 mg/l jasmonic acid), or (50, 100, 150, 200 mg/l salicylic acid). Result showed that there were significant differences between the various treatment, the best concentration of jasmonic acid to stimulate and increase the production was 8 mg/l for S. nigrum. Moreover, Salicylic acid at 100 mg/l stimulated the production of secondary metabolites in callus culture of S. nigrum.

Simple Sequence Repeat (SSR) markers were used to investigate the variations between drought sensitive and drought tolerant genotypes. Patterns obtained using RM328 and RM302 detected polymorphism between T14, T15 and Amber33, Amber Baghdad genotypes and those obtained using RM316 and RM201 for the tested genotypes suggested that these primers may have the ability to produce drought tolerance markers. According to the patterns obtained using RM189, RM3825 and RM212 primers indicated that these primers cannot be relied on as markers for drought tolerance.

Transgenic Sesbania grandiflora (L.) pers (Fabaceae) and Arabidopsis thaliana (L.) (Brassicaceae) plants harbouring the rabbit cytochrome p450 2E1 enzyme were evaluated for their ability to accumulate heavy metals, potassium (K), calcium (Ca), manganese (Mn), zinc (Zn), copper (Cu), iron (Fe), lead (Pb) and bromine (Br), using X-ray Fluorescence analysis. When grown for 15 days on heavy metal-contaminated soils, transgenic cuttings of S. grandiflora and T3 A. thaliana plants recorded higher dry and fresh weight compared with their respective controls (A. thaliana and S. grandiflora plants transformed with an empty vector). Dry weight of transgenic S. grandiflora plants (0.321 g) was seven times higher than that of the wildtype (0.049 g), and the fresh weight (4.421 g) was about 4.6 times higher. Likewise, the dry weight of CYP450 2E1 A. thaliana (0.198 g) was more than eight times higher than that seen in the control (0.024 g). Moreover, Fe, Mn, K, and Ca concentrations in transgenic plants were significa...

Bean pod mottle virus (BPMV) is a bipartite, positive-sense (+) RNA virus of Secoviridae . We recently reported that a 137 nucleotide (nt) stretch (#263–399) of the 466 nt 5′ untranslated region (5′ UTR) of BPMV RNA2 can be deleted without compromising BPMV propagation in host plants [Lin et al., J. Gen. Virol. 94 (2013) 1415–1420]. Here we demonstrate that nonviral insertions of up to 625 nt is tolerated by the same region. Furthermore, one insertion mutant underwent recombination in infected plants, leading to the truncation of nt #250–361, thus extending the dispensable sequence to 150 nt (nt #250–399). We are unaware of any other (+) RNA virus that tolerates insertion/deletion of these sizes (625 nt/150 nt) within its 5′ UTR. Importantly, tolerance of large insertions within the RNA2 5′ UTR offers a novel, more convenient site for incorporating host gene fragments, making BPMV a more versatile vector of virus-induced gene silencing.

This study was designed to evaluate the antibacterial activity of water, ethanol and acetone Myristica fragrans seed extract tested against four bacteria species; two Gram-positive bacteria (Bacillus subtilis, and Staphylococcus aureus) and two Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa). The susceptibility of these different bacterial species toward the extracts of this plants seed was compared with each other and between crust and bulb and with selected antibiotic (cipropholexin) used as positive control. Results showed that ethanol and acetone extracts exhibited antibacterial activity against gram positive bacterial species only, the diameter of inhibition zone reached 25mm against Staph. aureus by crust extract. Crust extract showed antibacterial activity more than the pulp extract. It is concluded that this plant can be indispensible source for secondary metabolites.

The yield of phenolic acids; rosmarinic acid (RA), caffeic acid (CAF) and phenolic diterpenes; carnosic acid (CA), carnosol (CAR), rosmanol (ROL) from rosemary (Rosmarinus officinalis L.) was investigated. Aqueous extraction was conducted for leaf and callus tissues, the chemical analysis for extracts was carried out using high performance liquid chromatography (HPLC). Data showed that RA and CAF production was 4.5, 2.7 µg/ml respectively. CA and CAR production reached 3.3, 2.8 µg/ml when the fungal elicitor; Fusarium oxysporum was added at the concentration 2.0 mg/l (2×10 4 CFU/ml), except ROL which was found at high levels (4.3, 4.2 and 4.6 µg/ml) only from leaf extracts, untreated callus and when callus treated with 0.4 mg/l of CaCl2 respectively.

This study was carried out to induce variations in tissue cultures of tomato (Lycopersicon esculentum Mill.) by using physical (gamma radiation) and chemical (sodium azide) mutagens. Different plant growth regulators were tested for their potentials in callus induction, and shoot and root formation. Callus initiation on hypocotyl explants was achieved on Murashige and Skoog’s (MS) medium supplemented with 0.1 mg/l 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.5 mg/l 6benzyl-aminopurine (BAP). Shoot regeneration was performed using hypocotyl explants cultured on MS medium containing 0.8 mg/l indole-3-acetic acid (IAA) and 2.0 mg/l BAP. Rooting was achieved on MS medium supplemented with 0.4 mg/l IAA. Callus cultures and seedlings were irradiated with gamma radiation at six doses 5, 15, 25, 35, 45 or 55 mGy from 137 Cs source, or treated with sodium azide(SA) at four concentrations 0.5, 1.0, 2.0 or 3.0 mM for 1, 3 or 6 hours. Callus fresh weights were increased after irradiation with 25 mGy gamma rays and treatment with (0.5 or 3.0) mM SA for 1 hour. Shoot regeneration was observed in all irradiated hypocotyl explants. Shoot regeneration at (15 and 35) mGy was similar to the control. Root formation was observed on hypocotyls irradiated with low doses (5 or 15) mGy only, while SA inhibited shoot and root regeneration. Random amplified polymorphic DNA (RAPD) technique was performed todetect genetic mutation in extracted DNA samples. DNA was extracted from mother plant, callus tissue, irradiated calli with 15 or 45 mGy of gamma ray and calli treated with 0.5 and 3 mM SA. Twelve arbitrary 10-mer primers were used of which eight primers gave amplification products. The total number of amplification products was 27 bands, of which 22 bands were monomorphic bands which indicates the presence of genetic mutations. One of these primers, C-02, revealed polymorphism in callus tissues treated with 45 mGy of gamma radiation, 0.5 and 3.0 mM SA. Others primers, C-08 and C-09, revealed polymorphisms in irradiated callus tissues with 15 mGy gamma radiation. RAPD was found to be effective technique to detect genetic mutations in callus tissue.

Plant tissue culture techniques are used for the micropropagation of Gladiolus, Oscar variety, using axillary buds of the flower stalks, axillary buds of corms and cormels. It was aimed to design production program for micropropagation and cormels production all year round. This work included many experiments. The effect of Benzyl adenine (BA) and Kinetin (Kin) and the interaction between BA and Naphthalene acetic acid (NAA) on shoot proliferation, the effect of BA on shoot multiplication, the effect of NAA on rooting after 30, 40 and 50 days, were studied. The role of the composed (Peatmoss only, Peatmoss: sand, sand only) in plantlet survival during acclimatization was also studied. Results showed that BA supplemented to the medium at 2.0 mgl induced shoot proliferation from axillary buds of flower stalks, giving an average shoot height of 2.5 cm. while reached 7.4 cm. in shoots induced from axillary buds of corms on a combination of 0.05 mgl NAA and 0.4 mgl BA. In cormels, the best response occurred at concentration of 1.0 mgl Kin giving average shoot length of (6.5 cm.). Maximum average of shoot number and length at the multiplication stage occurred at 0.75 mgl BA in axillary buds taken from both flower stalks and corms, while at 0.5 mgl in cormels. NAA at 1.0 mgl achieved the highest response in root number and length for all studied explants. Rooting percent, number and root length increased over time. Cormels formed 50 days after rooting reaching 100% cormels formation on all plantlets regenerated from the different explants under investigation. Peatmoss was found the best among the three tested media in sustaining plantlets survival.

Liquorice plant is considered one of the important medicinal and economical plants. It is rich with many compounds, minerals, vitamins, and even plant hormones. This research is aimed to study the possibility of using callus tissue extracts as an alternative to plant growth regulators added to the culture media. A factorial experiment was implemented to find out the appropriate combination between 2, 4-D and BA for callus induction on Liquoricenode explants. It was found that a combination of 2 mg/l 2, 4-D with 2.5 mg/l BA is the best one for callus induction and maintenance using MS medium. Water and alcoholic extracts were prepared from callus tissue at concentrations (0, 2, 4, 6, 8 or 10) ml/l then added to culture medium as an alternative to plant growth regulators. The effect of these concentrations on growth and development of tissues and organs for some plants was studied using soya bean, potato and wheat plants for this purpose. Results showed that water extract induced shoot proliferation from potato single nodes. Both types of extracts increased soya bean callus fresh weight significantly. It was found also that water extract was more effective than alcoholic one in increasing vegetative and root parts in germinating wheat seeds.

Iraq is the birthplace of the date palm, and historically it was the domestication center of this crop. Moreover, for some years, Iraq was the largest producer of dates in the world. Many factors negatively have affected both the production and natural genetic diversity of the crop. However, efforts are being made by the Iraqi authorities and researchers alike to compensate for the serious damage the date palm sector has experienced over the past 30 years. New approaches have been introduced including biotechnology, grove management, pest control, and industrial practices. Production limitations have been diagnosed and constraints are on their way to be resolved. Date palm plantations are under stress from many biotic and abiotic factors including key insect pests like dubas bug, lesser date moth, trunk and stalk borers, and Old World date mite. Date palm diseases cause serious damages to date palm trees especially where stress factors are present such as palm weakness, soil salinity, high water table, borers, and tree aging. The use of plant tissue culture to support propagation by offshoots is necessary and started in the early 1980s. Both direct organogenesis and callus induction with subsequent asexual embryo formation protocols were achieved. Approximately, 600 date palm cultivars were grown in Iraq before 1980; however, currently their number is reduced to 500. Morphological traits have been used to describe genetic variation in these cultivars which are mainly related to the fruit, leaf, trunk, and other parts of the tree. DNA marker analysis in Iraqi date palm is at the developmental stage and began in 2000. Tree management and fruit handling improvements are required urgently for better quality production. Date trading has to be reassessed in Iraq in order to overcome the outmoded market chains.

This study was conducted to determine the activity of plant Sesbania rostrata and two isolate from arbuscular mycorrhizae fungi (A,B) as a bioremediation of soil polluted by cadmium and lead elements in north and south of Baghdad city. The results showed that the average of soil pollution by cadmium and lead elements in north of Baghdad was less than the average of soil pollution in the south of Baghdad which recorded 10.0, 9.0 mg/kg and 27.0, 25.0 mg/kg respectively. The plant Sesbania recorded ability to accumulate the lead element in shoot system 19.65 mg/kg and in root system 27.2 mg/kg and for cadmium element 19.6, 24.6 mg/kg in shoot and root respectively. The results showed that the isolate A from soil pollution is more effected than isolate B which isolated from uncontaminated soil.

Sesbania grandiflora (L.) pers (Fabaceae) and Arabidopsis thaliana (L.) (Brassicaceae) were genetically engineered to constitutively express the rabbit cytochrome p450 2E1 enzyme aiming at increasing their activity toward trichloroethylene (TCE) and dichlorodiphenyltrichloroethane (DDT) removal Successful generation of Sesbania and Arabidopsis transgenic plants was verified using p450 2E1 specific PCR and confirmed by western blot analysis. Gas chromatography (GC) analysis revealed that small cuttings of Sesbania and third generation (F3) Arabidopsis transgenic plants exposed to TCE and DDT in small hydroponics' vessels accumulated more TCE and DDT compared to plants transformed with the empty vector. Furthermore, both transgenic plants were more effective in breaking down TCE and DDT with a 2-fold increase in TCE metabolism. Two independent Arabidopsis lines showed that DDT was metabolized about 4-fold higher than that detected in non transformed plants. Similarly, S. grandiflora cuttings removed 51 to 90% of the added DDT compared with only 3% removal in controls transformed with the null vector. Notably, stability of rabbit cytochrome p450 2E1 was confirmed using third generation Arabidopsis plants that displayed higher potential for the removal of two important pollutants, TCE and DDT compared with the controls.

An in vitro selection method was developed for Coleus blumei to enhance salt tolerance of this amenity species. Leaf disc explants were incubated on a Murashige & Skoog medium containing benzylaminopurine, 2 mg l-1, and napthalene acetic acid, 1 mg l-1, which initiated both callus and plantlets from the explants. A large number of explants were incubated on this differentiating medium containing 90 mM NaCl, which inhibited over 90% of plantlet formation. Surviving plantlets. were grown to maturity, when apical cuttings were taken and propagated. Plants were also allowed to flower and set seed. Cuttings from the selected regenerated plants showed consistently better growth in the presence of NaCl than unselected cuttings. Seed progeny of selected plants also showed more vigorous growth in the presence and absence of NaCl than progeny from unselected plants. The in vitro selection was compared with the results of an earlier in vivo selection to assess the contribution from tissue culture derived somaclonal variation. Progeny from the in vitro selection showed a higher level of tolerance than progeny from the in vivo selection.

SummaryThe aim of the investigation was to examine by in vivo screening the potential for salt tolerance in a selected cultivar of the amenity annual, Coleus blumei. Seedlings were placed on a capillary mat soaked in salinized nutrient solution. An initial dose response over the range 0-120 mM NaCl showed that at 90 mM NaCl only 15% of seedlings survived. A large scale screen of seedlings at 90 mM produced a number of survivors of which about 2% grew normally. Cuttings from these survivors showed more tolerance than cuttings from unselected plants. Seed progeny from some of these surviving plants were vegetatively more vigorous in the presence of 90 mM NaCl than seed progeny from unselected plants. Using this simple technique it was possible to screen large numbers of seedlings for salt tolerance in this species. It is suggested that the technique could also be used to screen for a wide range of other characters.

SummaryThe amenity species Coleus blumei and Salvia splendens were irrigated with increasing concentrations of NaCl under glasshouse conditions. Salt was either added to the plants from the seedling stage onwards at one of the following concentrations, 0, 30, 60,120 or 180 mM NaCl up to six weeks (Expt. I), or, starting from 0 mM NaCl, the concentration was increased by 15 mM NaCl per week to a final concentration of 210 mM NaCl at 14 weeks (Expt. II). In Expt. I leaf number, leaf area, shoot and root weight progressively decreased in Coleus. However, Salvia root and shoot weights showed a pronounced fall at 30 mM NaCl and total leaf abscission occurred at NaCl concentrations above 60 mM NaCl within six weeks. Leaf thickness increased in both species over the sampling period. Where there was a gradual increase in NaCl concentrations (Expt. II), growth of Coleus and Salvia was much less inhibited at concentrations of NaCl found to be inhibitory in Expt. I. In both the concentration of Na+ in the roots incr...