Summary
This study was carried out in the laboratories and greenhouse of the Biology Department, College of Science, Salahaddin-Erbil University, and the laboratories of the Research Center at Erbil Polytechnic University, from April 21st, 2019 to July 26th, 2020. The study consisted of three experiments, two of them are factorial (2x3x3) experiments, and the third consisted of three nano technique experiments. All experiments were applied according to a completely randomized design (CRD) with four replications. The first experiment was conducted to determine the effects of soil moisture (SM: 100% and 60% of field capacity), foliar application of nitrogen (N: 100, 200, and 300 kg/ha), and magnesium (Mg: 0.0, 30, and 60 kg/ha) on vegetative growth (VG), some physiological characteristics (PC), and yield components (YC) of rosemary plants (Rosmarinus officinalis L.). The second experiment was conducted to determine the effects of light intensity (LI: 100% and 62.37%) of the normal sunlight, ascorbic acid (AA: 0.0, 100, and 200 ppm), and benzyl adenine (BA: 0.0, 100, and 200 ppm) on some VG, PC, and YC rosemary plants. The third experiment was applied to determine the effects of different concentrations of methanol extract (MeRLE), ethyl acetate extract (EARLE), and rosemary essential oil (RMEO) from rosemary plant leaves using silver nanoparticle size (Ag-NPs) on Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Two cuttings were taken from rosemary shoots (in March, 2020 and July, 2020) after 11 and 14 months of planting, respectively.
Results of experiment 1 showed that: SM2 showed significant increases leaves moisture in cutting two (cut.2), proline in dry leaves, total carbohydrates, and relative yield of nitrogen in dry weight of shoots for cut.1 and cut.1 + 2, while SM2 decreased plant height, number of branches per plant, fresh weight (FW) and DW of shoots, and DW of leaves in cut.1, stem diameter cut.1, shoots dry weight in cut.2, FW and DW of stems cut.2, dry matter % (DM%) in leaves cut.2, phenolic compounds in leaves cut.1, relative yield in shoots DW cut.1,2,1+2, RMEO and concrete percentage, RMEO and concrete content per plant in DW of leaves cut.2. N2 increased the leaves FW cut.2, stem diameter in cut.2, and concrete weight for the DW cut.2. N3 registered the highest values in plant height, number of branches per plant, DW of shoots, leaves, and stems cut.1, FW of shoots cut1, DW of shoots, leaves, and stems cut.2, DM% in leaves and shoots, proline, phenolic compounds in leaves cut.2, Mean productivity (MP), Stress Tolerance Index (STI), RMEO % and in leaves dry weight cut.2 compared to N1. N3 decreased FW of leaves cut.2, stem diameter cut.2, moisture% in leaves and shoots cut.2 and relative decrease in yield (RDY). Mg2 increased roots FW and DW cut.2, chl.b in FW of leaves, root surface area (RSA), and root length density (RLD). Mg2 recorded significant decreases in FW of stems, RMEO%, and in dry weight of leaves cut.2. SM1N2 increased total carbohydrates% in DW of leaves cut.1, and RTD. SM1N3 increased the FW and DW of shoots in cut.1, DW of leaves and stems cut.1, shoots and leaves DW cut.2, DW of stems, DM% in leaves and shoots cut.2, phenolic compounds, RMEO and concrete% and leaves dry weight (L.DW) cut.2. SM2N2 increased stem diameter cut.2, FW of leaves cut.2, leaves moisture% in cut.2 significantly. SM2N3 decreased stem diameter, relative yield, N in shoots cut.2 and Mn. SM2N3 increased proline and RY in cut.1, 1+2. SM1Mg2 increased number of branches, FW of leaves, moisture% in leaves cut.2, RE of N in shoots DW cut.1+2, RMEO% and DW of leaves, and concrete% compared to SM1Mg1. SM1Mg3 increased the FW and DW of roots cut.2, moisture% in leaves cut.2, RSA, RMD, DRMD, RTD and RLD. N3Mg1 increased plant height, number of branches per plant, FW and DW of shoots cut.1, DW of leaves and stems cut.1, DW of shoots cut.2, FW and DW of stems cut.2, DM% in leaves cut.2, proline, phenolic compounds, MSTIk2, MP and RMEO% in leaves DW cut.2. SM2N1Mg1 decreased plant height, the number of branches, FW and DW of shoots cut.1, DW of leaves and stems cut.1, stem diameter cut.1, phenolic compounds and RTD. The triple interaction treatment SM1N3Mg3 increased the FW and DW of shoots cut.1, DW of leaves and stems cut.1, stem diameter cut.1, DM% in shoots cut.2 and phenolic compounds. SM1N3Mg3 decreased total carbohydrates% in dry weight of leaves cut.1, while SM1N3Mg2 increased RMEO and concrete (% and in g per plant) in DW of leaves cut.2.
Results of the experiment 2 showed that: LI2 increased plant height, DW of shoots and stems in cut.2, DW of stems cut.1+2, total plant leaf area (cm2), moisture% in leaves and shoots cut.2 and protein content in leaves DW cut.1. LI2 decreased chl.a, chl.b, and total chl.s in leaves fresh weight (L.FW), chl.a and car+ xan in L.DW, DW of shoots and leaves cut.1, DW of leaves and roots cut.2, DW of shoots and leaves cut.1+2, DW of whole plant and biological yield cut.2, specific leaf weight (SLW), roots diameter, roots volume, RSA, RMD, DRMD, RLD, DM% in leaves and shoots cut.2 and phenolic compounds in DW leaves. AA2 increased chl.b in FW of leaves, and MSTIk1. AA2 decreased car+xan in L.DW cut.1, L.DW cut.2 and RMEO in dry leaves cut.2. AA3 increased chl.a, chl.b, total chl.s in L.FW and DW, car+xan in dry leaves and L.DW cut.2. BA2 increased moisture% in leaves and shoots cut.2, Stress Susceptibility Index (SSI), Tolerance index (TOL), phenolic compounds in L.DW. BA3 increased chl.a, chl.b, total chl.s in L.FW, SLW, SSI, TOL and DM% in leaves and shoots cut.2 and RMEO cut.2. LI2AA3 increased chl.b, total chl.s in FW leaves, chl.a, chl.b, total chl.s in L.DW and moisture% in shoots cut.2. LI2AA1 decreased chl.a, chl.b, total chl.s in L.FW and DW, DW in shoots and leaves cut.1, DW of roots cut.2, DRMD and DM% in leaves and shoots cut.2. LI2AA1 significantly increased total PLA, moisture% in leaves and shoots cut.2, and RMEO cut.2. LI2BA2 decreased chl.a, chl.b, total chl.s in FW of leaves, chl.a, total chl.s and car+xan in L.DW, DW of shoots, leaves and roots cut.2, DW of shoots and leaves cut.1+2, DW of whole plant and biological yield cut.2, SLW, and shoots cut.2, RMEO cut.2. AA3BA2 increased chl.a, chl.b, total chl.s in L.FW and L.DW, car+xan in L.DW, DW of leaves cut.2 and SSI. AA1BA2 reduced chl.a, chl.b in L.FW, chl.b in L.DW, SLW, DM% in leaves and shoots cut.2, DI and protein content in L.DW cut.1. LI2AA3BA2 increased chl.a, chl.b, total chl.s in L.FW and L.DW, and root density. LI2AA3BA3 increased RMEO in L.DW cut.2. LI1AA1BA2 and LI1AA2BA2 increased phenolic compounds significantly. LI1AA3BA2 increased DW of shoots, leaves and stems cut.1, shoots and leaves in cut.2 and cut.1+2, roots dry weight (R.DW) cut.2, DW of whole plant, BY cut.2 and DRMD. LI2AA1BA1, LI2AA1BA3 and LI2AA3BA3 increased RMEO in dry leaves cut.2 significantly.
Additionally the results of the experiment 3 showed that the minimal inhibitory concentration (MIC) of methanol rosemary leaves extract (MeRLE) against E. coli was 300µg/ml without Ag-NPs and there were no effects on S. aureus and P. aeruginosa but the MIC of (MeRLE) with Ag-NPs against E. coli, S. aureus and P. aeruginosa was 300µg/ml. The MIC of (RMEO) against E. coli and S. aureus was 100µl/ml with or without Ag-NPs, and no effects of RMEO on P. aeruginosa were observed. MIC of ethyl acetate rosemary leaf extract (EARLE) against S. aureus with and without using Ag-NPs was 50µg/ml with inhibition zones of 15.2 and 12.5 mm, respectively. The MIC of EARLE against P. aeruginosa was 50µg/ml when used with nanoparticles only by 9.7 mm, and was 100µg/ml against E. coli with inhibition zones of 15.2 mm when used with Ag-NPs.
