Nghiên cứu xây dựng quy trình nhân giống in vitro lan rừng giả hạc xuân dendrobium anosmum lindl

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 NGHIÊN CỨU XÂY DỰNG QUY TRÌNH NHÂN GIỐNG IN VITRO LAN RỪNG GIẢ HẠC XUÂN (Dendrobium anosmum Lindl.) ThS Vương Thị Hồng Loan 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) NGHIÊN CỨU XÂY DỰNG QUY TRÌNH NHÂN GIỐNG IN VITRO LAN RỪNG GIẢ HẠC XUÂN (Dendrobium anosmum Lindl.) CƠ QUAN CHỦ TRÌ (Ký tên, đóng dấu xác nhận) CHỦ NHIỆM ĐỀ TÀI (Ký tên) Thành phố Hồ Chí Minh, tháng 01/2018 (Dendrobium anosmum Lindl.) lồi có hoa , in vitro + 25 g/L sucrose + g/L in vitro agar + 25 g/L RITA® sucrose in Hyponex® vitro Dendrobium anosmum Lindl BM20-QT.QLKH Trang ii Trang iii v vi vii 6 10 20 20 20 20 (Dendrobium anosmum Lindl.) 20 Danh sác Danh sác Thông tin 1.1 1.2 u v 2.1 2.2 2.3 2.3.1 2.3.1.1 in vitro 20 in 2.3.1.2 vitro 22 (Dendrobium anosmum 23 2.3.2 2.3.2.1 23 2.3.2.2 24 Dendrobium 24 25 26 28 (Dendrobium anosmum Lindl.) 28 in vitro in 28 2.3.3 anosmum Lindl.) in vitro 2.3.4 2.4 3.1 3.1.1 3.1.2 vitro 33 (Dendrobium anosmum 35 3.2 3.2.1 35 3.2.2 3.3 BM20-QT.QLKH 40 Dendrobium Trang iii anosmum Lindl.) in vitro in vitro 43 43 3.4 4.2 Ki n ngh BM20-QT.QLKH 50 53 53 53 54 Trang iv Tên v ANOVA Analysis of varience BA Benzyladenine ctv iên IAA Indole-3-3acetic acid IBA Indole-3-butyric acid KC Knudson C MS NAA naphthalene acetic acid PLBs Protocorm like bodies TDZ Thidiazuron TN BM20-QT.QLKH m Trang v Tên Trang 21 2.1 xuân in vitro in 2.2 vitro 22 xuân 2.3 23 xuân 2.4 24 xuân 2.5 25 xuân in vitro 3.1 28 xuân in vitro in vitro 3.2 33 xuân 3.3 35 xuân 3.4 36 xuân 3.5 41 xuân 3.6 43 xuân in vitro 3.7 BM20-QT.QLKH 50 Trang vi DANH SÁCH HÌNH Hình 2.1 2.2 3.1 Tên hình Lan xuân (Dendrobium anosmum Lindl ) in vitro (cao -2,5 cm) dù xuân in vitro in vitro 3.2 Trang 21 22 32 34 xuân 3.3 3.4 ng c a n BA NAA k t h p lên s PLBs tái sinh ch i t PLBs lan r ng Gi h c xuân ng c a ki u nuôi c y lên s sinh ch i t PLBs lan r ng Gi h c xuân 3.5 40 43 49 xuân in vitro 3.6 3.7 Cây xuân Cây hoa lan Thùy Anh) BM20-QT.QLKH 51 52 Trang vii in vitro xuân (Dendrobium anosmum Lindl.) Chuyên ngành: CNSH CM 28) 38862726 83904599 Fax: (08) 37990500 E-mail: hongloanvuong@gmail.com - Trung tâm Nghiên - 28 38862726 - Fax: 028 37990500 - Email: nghiencuu.ahtp@tphcm.gov.vn - Website: www.chta.com.vn 146.311.500 : a tá háng 12/2017 b in vitro xuân (Dendrobium anosmum in vitro xuân (Dendrobium anosmum Lindl.) xuân in vitro nh in vitro xuân xuân (Dendrobium anosmum Lindl.) xuân xuân xuân (Dendrobium anosmum Lindl.) in vitro xuân in vitro : in vitro la - xuân Dendrobium anosmum -80 % cm, x xuân in vitro UNIANOVA TyletaoPLBs BY BA TDZ /METHOD=SSTYPE(3) /INTERCEPT=INCLUDE /POSTHOC=BA TDZ(TUKEY) /PLOT=PROFILE(BA*TDZ) /EMMEANS=TABLES(BA) /EMMEANS=TABLES(TDZ) /EMMEANS=TABLES(BA*TDZ) /PRINT=ETASQ HOMOGENEITY DESCRIPTIVE /CRITERIA=ALPHA(.05) /DESIGN=BA TDZ BA*TDZ Univariate Analysis of Variance Between-Subjects Factors N BA TDZ 00 15 50 15 1.00 15 1.50 15 2.00 15 00 15 .05 15 10 15 20 15 50 15 Descriptive Statistics Dependent Variable:TyletaoPLBs BA TDZ Mean 00 00 17.7767 3.85093 05 33.3300 00000 10 42.2233 3.85093 20 44.4467 3.85093 50 48.8867 7.69608 Total 37.3327 12.03234 15 00 24.4467 3.85093 05 46.6667 6.66500 10 55.5533 3.85093 20 60.0000 6.67000 50 66.6700 00000 Total 50.6673 15.69554 15 00 33.3333 6.66500 05 62.2233 3.85093 10 96.2200 3.42291 20 97.7767 3.85093 50 86.6667 6.66500 Total 75.2440 25.73649 15 00 48.8900 3.84515 05 75.5533 3.85093 10 88.8900 3.84515 20 86.6667 6.66500 50 95.5533 3.85093 Total 79.1107 17.43232 15 00 53.3333 6.66500 05 71.1133 7.69608 10 91.1133 7.69608 20 68.8900 3.84515 50 57.7767 13.87510 Total 68.4453 15.42357 15 00 35.5560 14.83589 15 05 57.7773 16.84124 15 50 1.00 1.50 2.00 Total Std Deviation N .10 74.8000 22.82888 15 20 71.5560 20.07303 15 50 71.1107 19.29945 15 Total 62.1600 23.54731 75 a Levene's Test of Equality of Error Variances Dependent Variable:TyletaoPLBs F df1 1.892 df2 24 Sig 50 029 Tests the null hypothesis that the error variance of the dependent variable is equal across groups a Design: Intercept + BA + TDZ + BA * TDZ Tests of Between-Subjects Effects Dependent Variable:TyletaoPLBs Type III Sum of Source Partial Eta Squares df Mean Square F Sig Squared Corrected Model 39378.531a 24 1640.772 49.639 000 960 Intercept 289789.920 289789.920 8767.208 000 994 BA 18697.492 4674.373 141.417 000 919 TDZ 15827.241 3956.810 119.708 000 905 BA * TDZ 4853.798 16 303.362 9.178 000 746 Error 1652.692 50 33.054 Total 330821.143 75 41031.223 74 Corrected Total a R Squared = 960 (Adjusted R Squared = 940) Descriptive Statistics Dependent Variable:SoPLBstrungbinh BA TDZ 00 00 1.2200 19053 05 2.3300 00000 10 2.2200 19053 20 2.6667 33501 50 3.1133 50954 Total 2.3100 69675 15 00 2.1100 38105 05 3.7767 1.07221 10 4.4433 83578 50 Mean Std Deviation N 1.00 1.50 2.00 Total 20 4.1133 50954 50 4.0000 67000 Total 3.6887 1.05031 15 00 2.0000 33000 05 5.2233 50846 10 10.3300 2.64575 20 13.0000 1.85712 50 11.0000 1.19954 Total 8.3107 4.41167 15 00 3.4433 50954 05 10.3367 1.15470 10 11.3333 2.08167 20 10.6667 1.52753 50 10.6667 87877 Total 9.2893 3.24915 15 00 3.6667 57735 05 5.4433 1.01908 10 11.8867 50954 20 7.2233 50846 50 6.3300 00000 Total 6.9100 2.89577 15 00 2.4880 1.02257 15 05 5.4220 2.89019 15 10 8.0427 4.29070 15 20 7.5340 4.12625 15 50 7.0220 3.46236 15 Total 6.1017 3.85255 75 Levene's Test of Equality of Error Variancesa Dependent Variable:SoPLBstrungbinh F 3.903 df1 df2 24 Sig 50 000 Tests the null hypothesis that the error variance of the dependent variable is equal across groups a Design: Intercept + BA + TDZ + BA * TDZ Tests of Between-Subjects Effects Dependent Variable:SoPLBstrungbinh Type III Sum of Source Squares Partial Eta df Mean Square F Sig Squared 1045.671a 24 43.570 41.379 000 952 2792.336 2792.336 2651.942 000 981 BA 538.404 134.601 127.834 000 911 TDZ 302.799 75.700 71.894 000 852 BA * TDZ 204.467 16 12.779 12.137 000 795 Error 52.647 50 1.053 Total 3890.654 75 Corrected Total 1098.318 74 Corrected Model Intercept a R Squared = 952 (Adjusted R Squared = 929) SoPLBstrungbinh Subset BA Duncana,b N 00 15 50 15 2.00 15 1.00 15 1.50 15 Sig 2.3100 3.6887 6.9100 8.3107 9.2893 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) = 1.053 a Uses Harmonic Mean Sample Size = 15.000 b Alpha = 05 SoPLBstrungbinh Subset TDZ a,b Duncan N 00 15 05 15 50 15 7.0220 20 15 7.5340 10 15 Sig 2.4880 5.4220 7.5340 8.0427 1.000 Means for groups in homogeneous subsets are displayed Based on observed means The error term is Mean Square(Error) = 1.053 1.000 178 181 1.000 Tests of Between-Subjects Effects Dependent Variable:SoPLBstrungbinh Type III Sum of Source Partial Eta Squares df Mean Square F Sig Squared a 24 43.570 41.379 000 952 2792.336 2792.336 2651.942 000 981 BA 538.404 134.601 127.834 000 911 TDZ 302.799 75.700 71.894 000 852 BA * TDZ 204.467 16 12.779 12.137 000 795 Error 52.647 50 1.053 Total 3890.654 75 Corrected Total 1098.318 74 Corrected Model 1045.671 Intercept a Uses Harmonic Mean Sample Size = 15.000 b Alpha = 05 in vitro : xuân ONEWAY TylemautaoPLBs SoPLBstrungbinh BY Nghiemthuc /STATISTICS DESCRIPTIVES HOMOGENEITY /MISSING ANALYSIS /POSTHOC=DUNCAN ALPHA(0.05) Oneway Descriptives N TylemautaoPLBs Std Deviation Std Error 97.7767 3.85093 2.22333 69.4400 1.92518 1.11150 3 42.9700 2.04118 1.17847 28.0933 2.86088 1.65173 12 59.5700 27.85073 8.03981 13.0000 1.85712 1.07221 16.4333 1.00957 58288 3 9.9333 97275 56161 7.2267 39425 22762 12 11.6483 3.72688 1.07586 Total SoPLBstrungbinh Mean Total Descriptives 95% Confidence Interval for Mean Lower Bound TylemautaoPLBs Upper Bound Minimum Maximum 88.2104 107.3429 93.33 100.00 64.6576 74.2224 67.53 71.38 37.8994 48.0406 41.45 45.29 SoPLBstrungbinh 20.9865 35.2002 25.48 31.15 Total 41.8745 77.2655 25.48 100.00 8.3867 17.6133 11.00 14.67 13.9254 18.9412 15.27 17.08 7.5169 12.3498 9.27 11.05 6.2473 8.2060 6.87 7.65 Total 9.2804 14.0163 6.87 17.08 Test of Homogeneity of Variances Levene Statistic df1 df2 Sig TylemautaoPLBs 1.212 366 SoPLBstrungbinh 2.115 177 ANOVA Sum of Squares TylemautaoPLBs Between Groups Within Groups Total SoPLBstrungbinh Between Groups Within Groups Total df Mean Square 8470.520 2823.507 61.774 7.722 8532.294 11 141.647 47.216 11.140 1.392 152.786 11 TylemautaoPLBs a Duncan Subset for alpha = 0.05 Nghiemthuc N 3 3 Sig 28.0933 42.9700 69.4400 97.7767 1.000 1.000 1.000 1.000 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 SoPLBstrungbinh Duncana Subset for alpha = 0.05 Nghiemthuc N 3 3 3 7.2267 9.9333 13.0000 16.4333 F Sig 365.657 000 33.908 000 Sig 1.000 1.000 1.000 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 xuân UNIANOVA TangsinhPLBs BY BA NAA /METHOD=SSTYPE(3) /INTERCEPT=INCLUDE /POSTHOC=BA NAA(DUNCAN) /PLOT=PROFILE(BA*NAA) /PRINT=ETASQ HOMOGENEITY DESCRIPTIVE /CRITERIA=ALPHA(.05) /DESIGN=BA NAA BA*NAA Univariate Analysis of Variance Between-Subjects Factors N BA NAA 00 12 50 12 1.00 12 1.50 12 2.00 12 00 15 20 15 50 15 1.00 15 Descriptive Statistics Dependent Variable:TangsinhPLBs BA NAA 00 00 3.2833 40415 20 4.0833 86217 50 3.8167 12342 1.00 4.4133 32747 Total 3.8992 61135 12 00 5.3467 35557 20 6.1233 16166 50 6.9100 18358 1.00 5.6500 44542 Total 6.0075 67059 12 50 Mean Std Deviation N 1.000 1.00 1.50 2.00 Total 00 8.7467 43432 20 12.4700 73899 50 11.5000 66551 1.00 9.3667 60103 Total 10.5208 1.67308 12 00 9.1133 17214 20 10.7433 43294 50 11.4833 18903 1.00 11.0500 08888 Total 10.5975 96115 12 00 12.1700 26153 20 12.8133 42028 50 13.6567 59919 1.00 13.9800 81627 Total 13.1550 88413 12 00 7.7320 3.22374 15 20 9.2467 3.67091 15 50 9.4733 3.72657 15 1.00 8.8920 3.65180 15 Total 8.8360 3.54684 60 Levene's Test of Equality of Error Variancesa Dependent Variable:TangsinhPLBs F df1 2.542 df2 19 Sig 40 006 Tests the null hypothesis that the error variance of the dependent variable is equal across groups a Design: Intercept + BA + NAA + BA * NAA Tests of Between-Subjects Effects Dependent Variable:TangsinhPLBs Type III Sum of Source Squares Partial Eta df Mean Square F Sig Squared Corrected Model 733.258a 19 38.593 172.145 000 988 Intercept 4684.494 4684.494 20895.506 000 998 683.617 170.904 762.330 000 987 NAA 26.952 8.984 40.074 000 750 BA * NAA 22.690 12 1.891 8.434 000 717 Error 8.967 40 224 Total 5426.720 60 BA Corrected Total 742.226 59 a R Squared = 988 (Adjusted R Squared = 982) TangsinhPLBs Duncana,b Subset BA N 00 12 50 12 1.00 12 10.5208 1.50 12 10.5975 2.00 12 Sig 3.8992 6.0075 13.1550 1.000 1.000 694 1.000 Means for groups in homogeneous subsets are displayed Based on observed means The error term is Mean Square(Error) = 224 a Uses Harmonic Mean Sample Size = 12.000 b Alpha = 05 NAA TangsinhPLBs a,b Duncan Subset NAA N 00 15 1.00 15 20 15 9.2467 50 15 9.4733 Sig 7.7320 8.8920 1.000 1.000 197 Means for groups in homogeneous subsets are displayed Based on observed means The error term is Mean Square(Error) = 224 a Uses Harmonic Mean Sample Size = 15.000 b Alpha = 05 xuân ONEWAY TangsinhPLBs Taisinhchoi BY Nghiemthuc /STATISTICS DESCRIPTIVES EFFECTS HOMOGENEITY /MISSING ANALYSIS /POSTHOC=DUNCAN ALPHA(0.05) Oneway Descriptives 95% Confidence Interval for Mean Lower Bound TangsinhPLBs 00 10.6343 14.3057 1.00 14.1876 17.6391 2.00 14.9636 19.5831 3.00 18.6865 19.6668 Total 14.5395 17.8772 Fixed Effects 15.7457 16.6710 Random Effects 11.7070 20.7097 00 30.8986 43.6547 1.00 7.6151 9.8316 2.00 9.6922 19.0211 3.00 23.9642 105.6491 Total 15.9330 46.6487 Fixed Effects 25.7150 36.8667 Random Effects -9.3316 71.9133 Model Taisinhchoi Upper Bound Model Descriptives BetweenComponent Minimum TangsinhPLBs Maximum 00 11.62 12.96 1.00 15.27 16.65 2.00 16.22 17.98 3.00 18.95 19.31 Total 11.62 19.31 Model Fixed Effects Random Effects Taisinhchoi 7.84145 00 35.27 40.17 1.00 8.26 9.15 2.00 12.27 15.91 3.00 50.27 82.65 Total 8.26 82.65 Model Variance Fixed Effects Random Effects 628.34699 Test of Homogeneity of Variances Levene Statistic df1 df2 Sig TangsinhPLBs 2.099 179 Taisinhchoi 5.824 021 ANOVA Sum of Squares TangsinhPLBs Between Groups 24.007 3.864 483 75.887 11 5865.598 1955.199 561.267 70.158 6426.865 11 Total Between Groups Within Groups Total Mean Square 72.022 Within Groups Taisinhchoi df TangsinhPLBs a Duncan Subset for alpha = 0.05 Nghiemthuc N 00 1.00 2.00 3.00 Sig 12.4700 15.9133 17.2733 19.1767 1.000 1.000 1.000 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 Taisinhchoi a Duncan Subset for alpha = 0.05 Nghiemthuc N 1.00 8.7233 2.00 14.3567 00 3.00 Sig 37.2767 64.8067 434 1.000 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 1.000 1.000 F Sig 49.700 000 27.868 000