Khảo sát ảnh hưởng của xử lý đột biến bằng ems ethyl mathane sulfonate và tia gamma co60 đến sự biến dị của cây hoa đồng tiền gerbera jamesonii in vitro

BAN QUẢN LÝ KHU NÔNG NGHIỆP CÔNG NGHỆ CAO TP HCM TRUNG TÂM NGHIÊN CỨU VÀ PHÁT TRIỂN NÔNG NGHIỆP CÔNG NGHỆ CAO BÁO CÁO NGHIỆM THU KHẢO SÁT ẢNH HƯỞNG CỦA XỬ LÝ ĐỘT BIẾN BẰNG EMS (ETHYL METHANE SULFONATE) VÀ TIA GAMMA CO60 ĐẾN SỰ BIẾN DỊ CỦA CÂY HOA ĐỒNG TIỀN (Gerbera jamesonii) IN VITRO ThS Vương Thị Hồng Loan CN Kha Nữ Tú Uyên Thành phố Hồ Chí Minh, tháng 01/2018 BAN QUẢN LÝ KHU NÔNG NGHIỆP CÔNG NGHỆ CAO TP HCM TRUNG TÂM NGHIÊN CỨU VÀ PHÁT TRIỂN NÔNG NGHIỆP CÔNG NGHỆ CAO BÁO CÁO NGHIỆM THU (Đã chỉnh sửa theo góp ý Hội đồng nghiệm thu) KHẢO SÁT ẢNH HƯỞNG CỦA XỬ LÝ ĐỘT BIẾN BẰNG EMS (ETHYL METHANE SULFONATE) VÀ TIA GAMMA CO60 ĐẾN SỰ BIẾN DỊ CỦA CÂY HOA ĐỒNG TIỀN (Gerbera jamesonii) IN VITRO CƠ QUAN CHỦ TRÌ (Ký tên, đóng dấu xác nhận) CHỦ NHIỆM ĐỀ TÀI (Ký tên) Thành phố Hồ Chí Minh, tháng 01/2018 (Gerbera jamesonii) H in vitro tia gamma Co60 ; EMS (Gerbera jamesonii) in vitro 0,25 IBA 0,5 mg/L in vitro gamma Co60 EMS in vitro Gerbera jamesonii BM20-QT.QLKH Trang ii Trang ii iii v vi vii 6 20 29 29 29 Thông 1.1 1.2 2.1 2.2 2.2.1 in vitro in vitro 29 2.2.1.1 29 30 2.2.1.2 in vitro Co60 31 in vitro 60 2.2.2.1 31 in vitro 2.2.2.2 32 EMS 2.2.2 2.2.3 ng c a gamma Co60 EMS lên s t o r ng c a in vitro 33 2.3 34 35 in vitro 3.1 ây in vitro 35 3.1.1 35 3.1.2 in vitro 37 60 3.2 in vitro 40 3.2.1 60 40 3.2.2 BM20-QT.QLKH Trang iii 3.3 4.2 EMS ch in vitro 45 T o bi n d in vitro hoàn ch nh 49 53 53 53 54 PH L C BM20-QT.QLKH Trang iv ANOVA BA ctv CV Ducan Analysis of varience Benzyladenine Coefficient of variation V EMS Gy IAA IBA MS NAA NST TDZ TN BM20-QT.QLKH Ethyl Methane Sulfonate Gray Indole-3-3acetic acid Indole-3-butyric acid naphthalene acetic acid Thidiazuron Trang v D 2.1 Trang 29 2.2 30 in vitro 60 2.3 31 in vitro 2.4 32 in vitro 3.1 35 in vitro 3.2 3.3 60 38 41 in vitro 3.4 46 in vitro sau 30 ngày 3.5 BM20-QT.QLKH in vitro 50 Trang vi DANH SÁCH HÌNH Hình 1.1 1.2 3.1 Tên hình Trang 19 37 in vitro 3.2 3.3 60 in vitro gamma Co60 3.4 3.5 39 43 45 48 in vitro 3.6 3.7 3.8 BM20-QT.QLKH in vitro in vitro 50 52 52 Trang vii Ethyl Methane (Gerbera Sulfonate) tia gamma Co60 jamesonii) in vitro Chuyên ngành: CNSH 28) 38862726 83904599 Fax: (08) 37990500 E-mail: hongloanvuong@gmail.com Ch nhi m H tên: Kha N Tú Uyên 1985 H c v : C nhân Gi i tính: N Chuyên ngành: Công ngh Sinh h c t h c v : 2009 Ch c v : Chuyên viên Trung tâm Nghiên c u Phát tri n NNCNC Tp.HCM a ch p 1, xã Ph n tho (08) 8862726 i, huy n C Chi, Tp HCM Fax: (08) 37990500 a ch nhà riêng: n tho i: 0938896226 - - E-mail: uyentunu85@gmail.com Trung tâm Nghiên 28 38862726 BM20-QT.QLKH Trang - Fax: 028 37990500 - Email: nghiencuu.ahtp@tphcm.gov.vn - Website: www.chta.com.vn 7 : M c tiêu c th : in vitro - 60 (Gerbera jamesonii) in vitro in vitro 60 EMS in vitro tia gamma Co60 (Gerbera jamesonii) - in vitro in vitro BM20-QT.QLKH Trang Hassan Abu-Qaoud (2004), Direct Regeneration in Cyclamen Persicum Mill Using Seedling Tissues, An-Najah Univ J Res (N Sc.), Vol 18(2) Imelda M., Deswina P., Hartati S., Estiati A., and Atmowidjojo S., (2000), Chemical Mutation By Ethyl Methane Sulfonate (EMS) For Bunchy Top Virus Resistance Imtiyaz, Nazki T., Siddique M.A.A., Rather Z.A., Nelofar (2014), Studies on 60Co gamma irradiation for inducing in vitro mutagenesis in gerbera, Indian Journal of Horticulture, 71(1), pp77-81 Jong Yi Fang Siguina Traore (2011), In Vitro Mutation Induction of Saintpaulia Using Ethyl Methanesulfonate Khaliqa Shabbir, Touqeer Ahmad, Ishfaq Ahmad Hafiz1, Azhar Hussain, Nadeem Akhtar Abbasi and Jamil Ahmad, In vitro regeneration of Gerbera jamesonii cv Sunglow, African Journal of Biotechnology, 11(42), pp 9975-9984 Malgorzata Zalewska, Alicja Tymoszuk, Natalia Miler (2011), New Chrysanthemum cultivars as a result of in vitro mutagenesis with the application of different explant types, Acta Sci Pol., Hortorum Cultus 10(2) 2011, 109-123 Masao Sugiyama, Hiroyuki Saito, Hiroyuki Ichida, Yoriko Hayashi, Hiromichi Ryuto, Nobuhisa Fukunishi, Teruhiko Terakawa, Tomoko Abe (2008), Biological effects of heavy-ion beam irradiation on Cyclamen, Plant Biotechnology 25, 101 104 10 Marek Gaj and Malgorzata D Gaj (1996), The high frequency of variegated forms after in vitro mutagenesis in saintpaulia ionantha Wendl 11 M Dhivya, S Balakrishanan and T Thangaselvabai (2015), Sensitivity and mutagenic effects of Ethyl Methane Sulphonate on the growth of Crossandra Infundibuliformis L Nees, International Journal of Agricultural Science and Research (IJASR) Vol 5, Issue 3, Jun 2015, 199-204 12 Minerva Ghani, Surinder Kumar and Manisha Thakur (2015), Induction of novel variants through physical and chemical mutagenesis in Barbeton daisy (Gerbera jamesonii Hook.), page 585-590 BM20-QT.QLKH Trang 56 13 Nazma Akter, M.I Hoque and R.H Sarker (2012), In vitro Propagation in Three Varieties of Gerbera (Gerbera jamesonii Bolus.) from Flower Bud and Flower Stalk Explants, Plant Tissue Cult & Biotech 22(2): 143-152 14 Nor Azlina Hasbullah, Rosna Mat Taha and Asmah Awal (2008), Growth optimization and organogenesis of Gerbera Jamesonii Bolux ex Hook f In vitro, Pakistan Journal of Biological Sciences 11(11), 1449-1454 15 Ridgman WJ., (1989), Applied mutation breeding for vegetatively propagated crops, pp.132 16 Rodrigo Rocha Latado, Alvis Hernan Adames & Augusto Tulmann Neto (2004), In vitro mutation of chrysanthemum (Dendranthema grandiflora Tzvelev) with ethylmethanesulphonate (EMS) in immature floral pedicels, Plant Cell, Tissue and Organ Culture, 77: 103 106 17 Seneviratne K.A.C.N and D.S.A Wijesundara (2007), First African Violets (Saintpaulia ionantha, H Wendl.) With a Changing Colour Pattern Induced by Mutation, American BM20-QT.QLKH Journal of Plant Physiology, 2: 233-236 Trang 57 T ONEWAY hesonhanchoi chieucaochoi BY nghiemthuc /STATISTICS DESCRIPTIVES EFFECTS HOMOGENEITY /MISSING ANALYSIS /POSTHOC=DUNCAN ALPHA(0.05) Oneway Descriptives N hesonhanchoi Mean Std Deviation 00 2.4300 37041 21385 1.00 9.3300 40731 23516 2.00 8.9700 1.67248 96561 3.00 8.3533 59744 34493 4.00 6.6733 36364 20995 5.00 6.5667 62324 35983 6.00 7.0500 11790 06807 7.00 6.2233 65577 37861 8.00 6.1467 41065 23709 9.00 3.0867 45764 26422 10.00 5.8600 22650 13077 11.00 5.8467 63948 36920 12.00 4.8233 48429 27960 Total 39 6.4123 1.80415 28890 65013 10410 Model Fixed Effects Random Effects chieucaochoi Std Error 49072 00 1.6833 20306 11724 1.00 1.8267 16503 09528 2.00 1.5433 12220 07055 3.00 1.5233 04726 02728 4.00 1.6400 10583 06110 5.00 1.6500 17000 09815 6.00 1.5267 10408 06009 7.00 1.7267 08021 04631 8.00 1.5367 13317 07688 9.00 1.6533 17156 09905 10.00 1.7700 05568 03215 11.00 1.7900 07211 04163 12.00 1.6600 16523 09539 Total 39 1.6562 14812 02372 BM20-QT.QLKH Model Fixed Effects 13178 02110 Random Effects 02858 Test of Homogeneity of Variances Levene Statistic df1 df2 Sig hesonhanchoi 3.891 12 26 002 chieucaochoi 1.102 12 26 399 ANOVA Sum of Squares hesonhanchoi Between Groups Mean Square F 112.699 12 9.392 10.989 26 423 123.689 38 Between Groups 382 12 032 Within Groups 452 26 017 Total 834 38 Within Groups Total chieucaochoi df Post Hoc Test Homogeneous Subset hesonhanchoi a Duncan Subset for alpha = 0.05 nghiemthuc N 9.00 00 12.00 11.00 5.8467 5.8467 10.00 5.8600 5.8600 8.00 6.1467 7.00 6.2233 5.00 6.5667 4.00 6.6733 6.00 7.0500 3.00 8.3533 2.00 8.9700 1.00 9.3300 Sig 3.0867 2.4300 4.8233 1.000 465 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 BM20-QT.QLKH 4.8233 075 057 093 Sig 22.220 000 1.834 095 chieucaochoi Duncana Subset for alpha = 0.05 nghiemthuc N 3.00 1.5233 6.00 1.5267 8.00 1.5367 1.5367 2.00 1.5433 1.5433 4.00 1.6400 1.6400 1.6400 5.00 1.6500 1.6500 1.6500 9.00 1.6533 1.6533 1.6533 12.00 1.6600 1.6600 1.6600 00 1.6833 1.6833 1.6833 7.00 1.7267 1.7267 1.7267 10.00 1.7700 1.7700 1.7700 11.00 1.7900 1.7900 1.00 Sig 1.8267 061 054 147 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 : ONEWAY tylecayrare sore chieudaire BY nghiemthuc /POSTHOC=DUNCAN ALPHA(0.05) Oneway Descriptives N tylecayrare Mean Std Deviation 00 34.4100 6.84501 3.95197 1.00 83.5367 5.73740 3.31249 2.00 100.0000 00000 00000 3.00 100.0000 00000 00000 4.00 94.4400 2.02012 1.16632 5.00 100.0000 00000 00000 6.00 100.0000 00000 00000 Total 21 87.4838 23.11921 5.04503 3.46106 75527 Model Fixed Effects Random Effects sore Std Error 9.13838 00 2.0267 14978 08647 1.00 2.3833 55012 31761 2.00 3.2333 23714 13691 BM20-QT.QLKH 3.00 3.7033 36116 20851 4.00 3.7467 27025 15603 5.00 3.9067 36254 20931 6.00 4.2800 43509 25120 Total 21 3.3257 84461 18431 35966 07848 Model Fixed Effects Random Effects chieudaire 31442 00 2.1500 21071 12166 1.00 3.3867 35275 20366 2.00 3.5933 37608 21713 3.00 3.1833 42063 24285 4.00 2.6100 42930 24786 5.00 3.3000 38354 22143 6.00 2.9933 18502 10682 Total 21 3.0310 55610 12135 34905 07617 Model Fixed Effects Random Effects 18855 Test of Homogeneity of Variances Levene Statistic df1 df2 Sig tylecayrare 4.253 14 012 sore 1.217 14 354 563 14 753 chieudaire ANOVA Sum of Squares tylecayrare Between Groups 1753.709 167.705 14 11.979 10689.959 20 12.456 2.076 1.811 14 129 14.267 20 Between Groups 4.479 747 Within Groups 1.706 14 122 Total 6.185 20 Total Between Groups Within Groups Total chieudaire Mean Square 10522.254 Within Groups sore df Post Hoc Tests Homogeneous Subsets tylecayrare BM20-QT.QLKH F Sig 146.399 000 16.049 000 6.128 003 Duncana Subset for alpha = 0.05 nghiemthuc N 00 1.00 4.00 94.4400 2.00 100.0000 3.00 100.0000 5.00 100.0000 6.00 100.0000 Sig 34.4100 83.5367 1.000 1.000 095 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 sore a Duncan Subset for alpha = 0.05 nghiemthuc N 00 2.0267 1.00 2.3833 2.00 3.2333 3.00 3.7033 3.7033 4.00 3.7467 3.7467 5.00 3.9067 3.9067 6.00 4.2800 Sig .245 052 090 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 chieudaire Duncana Subset for alpha = 0.05 nghiemthuc N 00 2.1500 4.00 2.6100 6.00 2.9933 2.9933 3.00 3.1833 3.1833 5.00 3.3000 1.00 3.3867 BM20-QT.QLKH 2.6100 2.00 3.5933 Sig .129 076 076 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 ONEWAY tylesong chieucaochoi hesonhanchoi tylechoibiendi BY nghiemthuc /STATISTICS DESCRIPTIVES HOMOGENEITY /MISSING ANALYSIS /POSTHOC=DUNCAN ALPHA(0.05) Oneway Descriptives N tylesong chieucaochoi hesonhanchoi tylechoibiendi Mean Std Deviation Std Error 00 100.0000 00000 00000 1.00 96.8333 1.28850 74392 2.00 86.3433 4.56256 2.63419 3.00 76.2833 5.33113 3.07793 4.00 65.9233 4.56684 2.63667 5.00 51.1333 88636 51174 6.00 42.7800 2.61960 1.51243 Total 21 74.1852 21.10841 4.60623 00 1.6833 20306 11724 1.00 1.6233 12662 07311 2.00 1.6933 05132 02963 3.00 1.4833 11372 06566 4.00 1.5733 13317 07688 5.00 1.4733 14572 08413 6.00 1.4433 23288 13445 Total 21 1.5676 16177 03530 00 2.4300 05292 03055 1.00 2.3800 17692 10214 2.00 2.1567 21008 12129 3.00 1.7500 26514 15308 4.00 1.4167 06506 03756 5.00 1.1567 07024 04055 6.00 1.0867 07095 04096 Total 21 1.7681 55236 12054 00 0000 00000 00000 1.00 5.5667 60103 34700 2.00 10.6167 92398 53346 BM20-QT.QLKH 3.00 28.3333 2.66973 1.54137 4.00 34.1967 3.96901 2.29151 5.00 36.2667 5.42229 3.13056 6.00 37.8467 6.96102 4.01895 Total 21 21.8324 15.46737 3.37526 Test of Homogeneity of Variances Levene Statistic df1 df2 Sig tylesong 3.000 14 042 chieucaochoi 1.752 14 181 hesonhanchoi 1.490 14 252 tylechoibiendi 3.088 14 039 ANOVA Sum of Squares tylesong Between Groups 1458.749 158.804 14 11.343 8911.298 20 Between Groups 191 032 Within Groups 332 14 024 Total 523 20 5.777 963 325 14 023 6.102 20 4580.887 763.481 203.905 14 14.565 4784.792 20 Total hesonhanchoi Between Groups Within Groups Total tylechoibiendi Mean Square 8752.493 Within Groups chieucaochoi df Between Groups Within Groups Total F Sig 128.602 000 1.344 302 41.414 000 52.420 000 Post Hoc Tests Homogeneous Subsets tylesong a Duncan Subset for alpha = 0.05 nghiemthuc N 6.00 5.00 4.00 3.00 2.00 BM20-QT.QLKH 42.7800 51.1333 65.9233 76.2833 86.3433 1.00 96.8333 00 100.0000 Sig 1.000 1.000 1.000 1.000 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 chieucaochoi Duncana Subset for alpha = 0.05 nghiemthuc N 6.00 1.4433 5.00 1.4733 3.00 1.4833 4.00 1.5733 1.00 1.6233 00 1.6833 2.00 1.6933 Sig .098 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 hesonhanchoi a Duncan Subset for alpha = 0.05 nghiemthuc N 6.00 1.0867 5.00 1.1567 4.00 3.00 2.00 2.1567 1.00 2.3800 00 2.4300 Sig 1.1567 1.4167 1.7500 583 056 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 BM20-QT.QLKH 1.000 055 1.000 269 tylechoibiendi Duncana Subset for alpha = 0.05 nghiemthuc N 00 0000 1.00 5.5667 2.00 3.00 28.3333 4.00 34.1967 5.00 36.2667 6.00 37.8467 5.5667 10.6167 Sig .096 127 081 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 UNIANOVA tylesong BY EMS Thoigian /DESIGN=EMS Thoigian EMS*Thoigian Univariate Analysis of Variance Between-Subjects Factors N EMS Thoigian 20 12 40 12 60 12 80 12 1.00 12 2.00 12 3.00 12 4.00 12 Descriptive Statistics Dependent Variable:tylesong EMS Thoigian Mean 20 1.00 85.8633 3.96310 2.00 80.4367 81002 3.00 72.1067 3.53543 4.00 63.2133 1.97014 Total 75.4050 9.28399 12 BM20-QT.QLKH Std Deviation N 34.1967 285 .40 60 80 Total 1.00 74.9333 4.73437 2.00 59.0900 6.36720 3.00 50.5533 1.43959 4.00 38.9833 3.33355 Total 55.8900 14.18672 12 1.00 37.4600 4.98771 2.00 33.8367 3.01027 3.00 30.6067 74272 4.00 28.7200 4.35039 Total 32.6558 4.66443 12 1.00 28.2600 3.38433 2.00 16.6500 1.19264 3.00 5.0333 91882 4.00 2.0000 1.76321 Total 12.9858 10.97779 12 1.00 56.6292 25.64573 12 2.00 47.5033 25.54149 12 3.00 39.5750 25.91789 12 4.00 33.2292 23.07561 12 Total 44.2342 25.82383 48 Levene's Test of Equality of Error Variancesa Dependent Variable:tylesong F df1 1.779 df2 15 Sig 32 084 Tests the null hypothesis that the error variance of the dependent variable is equal across groups a Design: Intercept + EMS + Thoigian + EMS * Thoigian Tests of Between-Subjects Effects Dependent Variable:tylesong Type III Sum of Source Squares Partial Eta df Mean Square F Sig Squared 30984.652a 15 2065.643 184.505 000 989 Intercept 93919.752 93919.752 8389.010 000 996 EMS 26615.945 8871.982 792.455 000 987 3685.696 1228.565 109.737 000 911 EMS * Thoigian 683.010 75.890 6.779 000 656 Error 358.258 32 11.196 Corrected Model Thoigian BM20-QT.QLKH Total Corrected Total 125262.662 48 31342.910 47 a R Squared = 989 (Adjusted R Squared = 983) Estimated Marginal Means EMS Dependent Variable:tylesong 95% Confidence Interval EMS Mean Std Error Lower Bound Upper Bound 20 75.405 966 73.438 77.372 40 55.890 966 53.923 57.857 60 32.656 966 30.688 34.623 80 12.986 966 11.018 14.953 Thoigian Dependent Variable:tylesong 95% Confidence Interval Thoigian Mean Std Error Lower Bound Upper Bound 1.00 56.629 966 54.662 58.597 2.00 47.503 966 45.536 49.471 3.00 39.575 966 37.608 41.542 4.00 33.229 966 31.262 35.197 EMS * Thoigian Dependent Variable:tylesong 95% Confidence Interval EMS Thoigian 20 1.00 85.863 1.932 81.928 89.798 2.00 80.437 1.932 76.502 84.372 3.00 72.107 1.932 68.172 76.042 4.00 63.213 1.932 59.278 67.148 1.00 74.933 1.932 70.998 78.868 2.00 59.090 1.932 55.155 63.025 3.00 50.553 1.932 46.618 54.488 4.00 38.983 1.932 35.048 42.918 1.00 37.460 1.932 33.525 41.395 2.00 33.837 1.932 29.902 37.772 3.00 30.607 1.932 26.672 34.542 4.00 28.720 1.932 24.785 32.655 1.00 28.260 1.932 24.325 32.195 40 60 80 BM20-QT.QLKH Mean Std Error Lower Bound Upper Bound 2.00 16.650 1.932 12.715 20.585 3.00 5.033 1.932 1.098 8.968 4.00 2.000 1.932 -1.935 5.935 Post Hoc Tests EMS Homogeneous Subsets tylesong a,b Duncan Subset EMS N 80 12 60 12 40 12 20 12 12.9858 32.6558 55.8900 75.4050 Sig 1.000 1.000 1.000 1.000 Means for groups in homogeneous subsets are displayed Based on observed means The error term is Mean Square(Error) = 11.196 a Uses Harmonic Mean Sample Size = 12.000 b Alpha = 05 Thoigian Homogeneous Subsets tylesong a,b Duncan Subset Thoigian N 4.00 12 3.00 12 2.00 12 1.00 12 Sig 33.2292 39.5750 47.5033 56.6292 1.000 1.000 1.000 Means for groups in homogeneous subsets are displayed Based on observed means The error term is Mean Square(Error) = 11.196 a Uses Harmonic Mean Sample Size = 12.000 b Alpha = 05 BM20-QT.QLKH 1.000 BM20-QT.QLKH

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