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Abstract [Objectives] This study was conducted to improve the efficiency of callus induction and redifferentiation， and construct highfrequency plant regeneration techniques of tissue culture in Anthuium andraeanum.
[Methods] The effects of different genotypes， explant types and hormonal conditions on callus induction and redifferentiation of A. andraeanum were studied by using the aseptic A. andraeanum testtube plantlets as test materials.
[Results] Among the four kinds of aseptic A. andraeanum plantlets， the callus induction using stem segments with leaves was the best， followed by stem segments and leaves， and the petioles were the worst; among the six A. andraeanum varieties tested， the callus production rates of four varieties reached 100%; and the callus differentiation rate reached 93.3%-100%through the organogenesis pathway， and the suitable differentiation medium was 1/2MS +ZT 0.5 mg/L + 2，4D 0.1 mg/L.
[Conclusions] The research results provide a new experimental basis for optimizing the technical system of A. andraeanum rapid propagation.
Key words Anthuium andraeanum; Explant type; Callus induction; Cytokinin; Adventitious bud
Anthuium andraeanum is a perennial herb in Araceae. It has elegant posture， and its flower leaves are both ornamental. In the global flower trade， its quantity of sale is the second largest tropical flower only second to tropical orchids[1]. The production of A. andraeanum is mainly through the organogenesis pathway which uses the leaves， petioles[4-5]and stem segments[5]of potted seedlings[2-3]as explants， but aseptic seedlings can also achieve good results as explants. For example， Du[6]used leaves， petioles and stem segments of aseptic seedlings as explants to perform invitro culture， achieving the callus induction rate higher than that of potted seedlings. Huang[7]used the aseptic seedlings of such four A. andraeanum varieties as ‘Tropic’， ‘Rapido’， ‘Atlanta’ and ‘Arizona’ to compare the culture effects of different parts of explants， and found that the stem segments showed the highest induction rate with shorter callus production time and differentiation time. It took only about 50 d to obtain complete plants with roots and leaves， but the effects of invitro culture between different varieties were also quite different. To this end， in this study， aseptic testtube plantlets of six A. andraeanum varieties were selected as experimental materials to study the effects of different genotypes， explant types and hormone conditions on callus induction and redifferentiation， in order to further optimize the cultivation of A. andraeanum. This study provides a new experimental basis for traditional technology. Materials and Methods
Plant materials
The test materials were aseptic testtube plantlets of A. andraeanum ‘Alabama’， ‘Turenza’， ‘Pink champion’， ‘Madural’， ‘Yalihong’ and ‘Hongchun’， which were subcultured at （25±2） ℃ under the conditions of 1 500-2 000 lx light and 12 hlight and dark once every 30 d.
Test division
Comparative test of different types of explants
With the A. andraeanum ‘Alabama’， the leaves， petioles， stem segments and stem segments with leaves were harvested as explants， and the treatment methods were as follows. Leaves： After removing the petioles and scratching veins at the leaf base， the leaves were inoculated in the medium with the back downward. Petioles： The petioles were cut into sections with a length of 0.5 cm. Stem segments： The leaves and petioles were removed， and the stems were cut into segments with a length of 1 cm. Stem segments with leaves： The excess leaves and adventitious roots of the aseptic plantlets were removed， the growth points and two young leaves were retained at the top， and the stem segments of about 1 cm in length were cut and inserted vertically into the callus induction medium. Each treatment was repeated 3 times with 10 explants per time.
Comparative test of different varieties
With aseptic plantlets of A. andraeanum ‘Alabama’， ‘Turenza’， ‘Pink champion’， ‘Madural’， ‘Yalihong’ and ‘Hongchun’ as the test materials， the stem segments with leaves were inoculated into the callus induction medium. Each variety was repeated 4 times with 10 explants per time.
Comparative test of cytokinin types and concentrations
The calli of A. andraeanum ‘Alabama’ were selected as the test material. They were cut into small pieces of about 0.5 cm×0.5 cm×1 cm as explants， and inoculated into the callus differentiation medium containing different cytokinins. The cytokinin conditions were 6BA 1.0 mg/L （denoted as MSB1）， 6BA 0.5 mg/L （MSB2）， ZT 1.0 mg/L （MSZ1）， ZT 0.5 mg/L （MSZ2）， TDZ 0.1 mg/L （MST1） and TDZ 0.05 mg/L （MST2）. Each treatment was repeated 3 times with 10 explants per time.
Investigation items
Subculture was performed once every 30 d. After 60 d of inoculation， the callus production rate， callus size， differentiation degree and adventitious bud morphology were investigated. Specifically， the calculation was performed according Callus size = The long diameter of the irregular callus， Callus production rate = The number of explants forming callus/The number of inoculated explants × 100%， Relative growth of callus = The difference between the initial value and the final value of the long diameter of the callus piece， and The adventitious bud differentiation rate = The number of differentiated callus pieces/The number of inoculated callus pieces × 100%. The data were initially collated by EXCEL， and statistical analysis was performed using SPSS18.0 analysissoftware. Results and Analysis
Effects of different types of explants from A. andraeanum asepticplantlets on callus induction
In order to investigate the effect of explants on the callus induction of A. andraeanum， the leaves， petioles， stem segments and stem segments with leaves from ‘Alabama’ aseptic plantlets were used as explants to induce calli in the callus induction medium. The results are shown in Table 1. Among the four explants， the callus induction rates of stem segments and stem segments with leaves were both 100%， while the callus induction rate of petioles was the lowest and had a significant difference from other materials. The stem segments with leaves showed the largest callus pieces， followed by the stem segments， and the petioles and leaves exhibited calli of about the same but low sizes， with a significant difference from other two materials.
The callus induction and growth status are shown in Fig. 1. With the stem segments and the stem segments with leaves as explants， a better callus induction effect was achieved， and the calli were compact， slightly green. The calli from the stem segments with leaves were produced from the base and were spherical， and at 60 d after inoculation， a large number of adventitious buds could be observed. The calli from the stem segments also showed a small amount of differentiated adventitious buds， while calli from the petioles and leaves were not differentiated， which meant that the callus induction effects were significantly inferior to the stem segments with leaves and the stem segments.
Effects of different A. andraeanum varieties on the callus induction from stem segments with leaves
On the basis of the above tests， the effects of different A. andraeanum varieties on the callus induction of stem segments with leaves were further explored. The results are shown in Table 2. The callus induction rates of the six varieties tested exceeded 70%. ‘Turenza’， ‘Pink champion’， ‘Yalihong’ and ‘Alabama’ had the same callus induction rate of 100%， while the induction rate of ‘Hongchun’ was the lowest at 70.17%. Secondly， from the size of the induced callus pieces， the order was ‘Pink champion’>‘Turenza’>‘Alabama’， ‘Yalihong’>‘Madural’， ‘Hongchun’.
From the callus morphology （Fig. 2）， the induced calli were green in color and dense in texture. The differentiation of adventitious buds was observed 60 d after inoculation. It can be considered that the stem segments with leaves from aseptic plantlets as explants can obtain better results for callus induction， and the method is suitable for most A. andraeanum varieties. Effects of cytokinin types and concentrations on callus redifferentiation of A. andraeanum
The callus of A. andraeanum ‘Alabama’ was used as experimental material to investigate the effects of different cytokinin types and concentrations on adventitious bud differentiation. The results are shown in Table 3. Under different hormonal conditions， the callus redifferentiation rate reached more than 90%， but there were significant differences in differentiation degree and adventitious bud morphology. Combined with Table 3 and Fig. 3， it can be seen that when the cytokinin concentration was high， the morphology of adventitious buds formed by redifferentiation was abnormal. Therefore， after primary culture and induction of callus， appropriate reduction of cytokinin concentration is conducive to adventitious bud differentiation， in which 6BA and ZT concentrations should not exceed 0.5 mg/L and TDZ concentration should not exceed 0.05 mg/L.
Xiuxiu LI et al. Study on Highfrequency Callus Induction From Aseptic Plantlets of Anthuium andraeanum
Discussion
In the research and application of A. andraeanum tissue culture and rapid propagation， the plants are mainly regenerated by the organogenesis pathway of callus， and most of the explants are from potted seedlings or aseptic seedlings[8-9]. Liu[10]induced calli using the leaves， petioles and roots of the aseptic seedlings of A. andraeanum ‘Pink champion’ as explants， and the average callus production rate of the leaves was 70.1%， which was higher than those of the petioles and root segments. However， in this study， the callus production rate of the leaves from the ‘Alabama’ aseptic plantlets was only 46.7%. The reason for this difference may be related to A. andraeanum varieties， culture media and explant treatment methods. The leaf explants were free of removal of leaf opex and had a small wound area， resulting in a lower callus production rate.
The callus induction effect of A. andraeanum is also affected by its genotype and explant treatment. For example， among the test varieties ‘Robino’， ‘Pink champion’， ‘Champion’， ‘ Sweetheart Red’， ‘Arizona’ and ‘Moli’， the callus induction rate of the ‘Champion’ aseptic seedlings can reach 96.7%， while ‘Moli’ failed to induce callus[10]. Such three varieties as ‘Kratt’， ‘Arizona’ and ‘Atlanta’ all showed the callus induction rates of stem segments with petioles reaching 63% or more[11]. In this study， when the callus induction was carried out with the stem segments as the explants， the four varieties， ‘Pink champion’， ‘Turenza’， ‘Alabama’ and ‘Yalihong’ all exhibited the callus production rate of 100%， and ‘Madural ’ and ‘Hongchun’ showed the callus production rates higher than 70%， indicating that the callus induction rate had been significantly improved， which might be related to their genotypes and the explant treatment method. In this study， the excised stem explants retained the upper growth point and two young leaves， suggesting that the combination of endogenous and exogenous phytohormones would facilitate the formation of callus. This study also showed that a high concentration of thidiazepine （TDZ） can cause the malformation of the differentiated adventitious buds， which is similar to the test results of Wang et al.[12]， that is， the induction rate of A. andraeanum callus increases with the concentration of TDZ increasing， but too high concentration is teratogenic to adventitious bud differentiation. In this study， when the concentration of TDZ was 2.0mg/L in the primary culture medium， various varieties still had a better callus induction effect， but after being transferred to the medium with the TDZ concentration of 0.05 mg/L， the adventitious buds were manifested as malformed plantlets. Therefore， in the process of inducing dedifferentiation and adventitious bud redifferentiation of A. andraeanum callus， the concentration of TDZ should not be too high.
References
[1] WU AL. Tissue culture in vitro and rapid propagation of Anthurium androaeanum[J]. Genomics and Applied Biology， 2010， 29（1）： 185-190. （in Chinese）
[2] DANG CJ， ZHOU ZQ， LIU C， et al. Tissue culture and reproduction of Anthurium varieties[J]. Journal of Agricultural Sciences， 2015， （1）： 61-64. （in Chinese）
[3] FINNIE JF， STADEN JV. Invitro culture of Anthurium andraeanum[J]. South African Journal of Botany， 1986， 52（4）：343-346.
[4] NIU RH. Construction of Anthuium andraeanum genetic transformation system and antibacterial peptide gene expression vector[D]. Suzhou： Suzhou University， 2015： 13-17. （in Chinese）
[5] CAI N. Optimization of largescale invitro culture techniques for Anthuium andraeanum[D]. changsha： agricultural university of Hunan， 2004： 14-21. （in Chinese）
[6] DU JR. Study on tissue culture technology of Anthuium andraeanum[D]. baoding： agricultural university of Hebei， 2010：14-23. （in Chinese）
[7] HUANG LF. Technical optimization on factory production of Anthuium andraeanum tissue culture seedlings[D]. changsha： agricultural university of Hunan， 2008： 15-23. （in Chinese）
[8] JIA Y， MA Y， GUO Y， et al. Study on tissue culture of Anthurium andraeanum Lind[J]. Journal of Henan Normal University， 2007， 35（1）：164-166.
[9] LAN QY， LI QR， HE HY， et al. The callus induction of Anthurium andraeanum Linden and bud differentiation[J]. Acta Horticulturae Sinica， 2003， （1）：107-109. （in Chinese）
[10] LIU BJ. Agrobacteriummediated Transformation of AtCBF3 and PaFT Genes into Anthurium andraeanum[D]. Wuhan： Huazhong Agricultural University， 2011： 14-21. （in Chinese）
[11] YAO Z. Genetic transformation of Anthurium andraeanum and Phalaenopsis aphrodite Rchb. F. by carotenoid synthase genes PSY， PDS， LycB and LycE[D]. Changchun： Jilin University， 2005： 23-28. （in Chinese）
[12] WANG Y， REN M， YANG Y. Application of thidiazuron on tissue culture in Anthurium andraeanum[J]. Tianjin Agricultural Sciences， 2013， 19（9）： 12-14. （in Chinese）
Editor： Yingzhi GUANG Proofreader： Xinxiu ZHU
[Methods] The effects of different genotypes， explant types and hormonal conditions on callus induction and redifferentiation of A. andraeanum were studied by using the aseptic A. andraeanum testtube plantlets as test materials.
[Results] Among the four kinds of aseptic A. andraeanum plantlets， the callus induction using stem segments with leaves was the best， followed by stem segments and leaves， and the petioles were the worst; among the six A. andraeanum varieties tested， the callus production rates of four varieties reached 100%; and the callus differentiation rate reached 93.3%-100%through the organogenesis pathway， and the suitable differentiation medium was 1/2MS +ZT 0.5 mg/L + 2，4D 0.1 mg/L.
[Conclusions] The research results provide a new experimental basis for optimizing the technical system of A. andraeanum rapid propagation.
Key words Anthuium andraeanum; Explant type; Callus induction; Cytokinin; Adventitious bud
Anthuium andraeanum is a perennial herb in Araceae. It has elegant posture， and its flower leaves are both ornamental. In the global flower trade， its quantity of sale is the second largest tropical flower only second to tropical orchids[1]. The production of A. andraeanum is mainly through the organogenesis pathway which uses the leaves， petioles[4-5]and stem segments[5]of potted seedlings[2-3]as explants， but aseptic seedlings can also achieve good results as explants. For example， Du[6]used leaves， petioles and stem segments of aseptic seedlings as explants to perform invitro culture， achieving the callus induction rate higher than that of potted seedlings. Huang[7]used the aseptic seedlings of such four A. andraeanum varieties as ‘Tropic’， ‘Rapido’， ‘Atlanta’ and ‘Arizona’ to compare the culture effects of different parts of explants， and found that the stem segments showed the highest induction rate with shorter callus production time and differentiation time. It took only about 50 d to obtain complete plants with roots and leaves， but the effects of invitro culture between different varieties were also quite different. To this end， in this study， aseptic testtube plantlets of six A. andraeanum varieties were selected as experimental materials to study the effects of different genotypes， explant types and hormone conditions on callus induction and redifferentiation， in order to further optimize the cultivation of A. andraeanum. This study provides a new experimental basis for traditional technology. Materials and Methods
Plant materials
The test materials were aseptic testtube plantlets of A. andraeanum ‘Alabama’， ‘Turenza’， ‘Pink champion’， ‘Madural’， ‘Yalihong’ and ‘Hongchun’， which were subcultured at （25±2） ℃ under the conditions of 1 500-2 000 lx light and 12 hlight and dark once every 30 d.
Test division
Comparative test of different types of explants
With the A. andraeanum ‘Alabama’， the leaves， petioles， stem segments and stem segments with leaves were harvested as explants， and the treatment methods were as follows. Leaves： After removing the petioles and scratching veins at the leaf base， the leaves were inoculated in the medium with the back downward. Petioles： The petioles were cut into sections with a length of 0.5 cm. Stem segments： The leaves and petioles were removed， and the stems were cut into segments with a length of 1 cm. Stem segments with leaves： The excess leaves and adventitious roots of the aseptic plantlets were removed， the growth points and two young leaves were retained at the top， and the stem segments of about 1 cm in length were cut and inserted vertically into the callus induction medium. Each treatment was repeated 3 times with 10 explants per time.
Comparative test of different varieties
With aseptic plantlets of A. andraeanum ‘Alabama’， ‘Turenza’， ‘Pink champion’， ‘Madural’， ‘Yalihong’ and ‘Hongchun’ as the test materials， the stem segments with leaves were inoculated into the callus induction medium. Each variety was repeated 4 times with 10 explants per time.
Comparative test of cytokinin types and concentrations
The calli of A. andraeanum ‘Alabama’ were selected as the test material. They were cut into small pieces of about 0.5 cm×0.5 cm×1 cm as explants， and inoculated into the callus differentiation medium containing different cytokinins. The cytokinin conditions were 6BA 1.0 mg/L （denoted as MSB1）， 6BA 0.5 mg/L （MSB2）， ZT 1.0 mg/L （MSZ1）， ZT 0.5 mg/L （MSZ2）， TDZ 0.1 mg/L （MST1） and TDZ 0.05 mg/L （MST2）. Each treatment was repeated 3 times with 10 explants per time.
Investigation items
Subculture was performed once every 30 d. After 60 d of inoculation， the callus production rate， callus size， differentiation degree and adventitious bud morphology were investigated. Specifically， the calculation was performed according Callus size = The long diameter of the irregular callus， Callus production rate = The number of explants forming callus/The number of inoculated explants × 100%， Relative growth of callus = The difference between the initial value and the final value of the long diameter of the callus piece， and The adventitious bud differentiation rate = The number of differentiated callus pieces/The number of inoculated callus pieces × 100%. The data were initially collated by EXCEL， and statistical analysis was performed using SPSS18.0 analysissoftware. Results and Analysis
Effects of different types of explants from A. andraeanum asepticplantlets on callus induction
In order to investigate the effect of explants on the callus induction of A. andraeanum， the leaves， petioles， stem segments and stem segments with leaves from ‘Alabama’ aseptic plantlets were used as explants to induce calli in the callus induction medium. The results are shown in Table 1. Among the four explants， the callus induction rates of stem segments and stem segments with leaves were both 100%， while the callus induction rate of petioles was the lowest and had a significant difference from other materials. The stem segments with leaves showed the largest callus pieces， followed by the stem segments， and the petioles and leaves exhibited calli of about the same but low sizes， with a significant difference from other two materials.
The callus induction and growth status are shown in Fig. 1. With the stem segments and the stem segments with leaves as explants， a better callus induction effect was achieved， and the calli were compact， slightly green. The calli from the stem segments with leaves were produced from the base and were spherical， and at 60 d after inoculation， a large number of adventitious buds could be observed. The calli from the stem segments also showed a small amount of differentiated adventitious buds， while calli from the petioles and leaves were not differentiated， which meant that the callus induction effects were significantly inferior to the stem segments with leaves and the stem segments.
Effects of different A. andraeanum varieties on the callus induction from stem segments with leaves
On the basis of the above tests， the effects of different A. andraeanum varieties on the callus induction of stem segments with leaves were further explored. The results are shown in Table 2. The callus induction rates of the six varieties tested exceeded 70%. ‘Turenza’， ‘Pink champion’， ‘Yalihong’ and ‘Alabama’ had the same callus induction rate of 100%， while the induction rate of ‘Hongchun’ was the lowest at 70.17%. Secondly， from the size of the induced callus pieces， the order was ‘Pink champion’>‘Turenza’>‘Alabama’， ‘Yalihong’>‘Madural’， ‘Hongchun’.
From the callus morphology （Fig. 2）， the induced calli were green in color and dense in texture. The differentiation of adventitious buds was observed 60 d after inoculation. It can be considered that the stem segments with leaves from aseptic plantlets as explants can obtain better results for callus induction， and the method is suitable for most A. andraeanum varieties. Effects of cytokinin types and concentrations on callus redifferentiation of A. andraeanum
The callus of A. andraeanum ‘Alabama’ was used as experimental material to investigate the effects of different cytokinin types and concentrations on adventitious bud differentiation. The results are shown in Table 3. Under different hormonal conditions， the callus redifferentiation rate reached more than 90%， but there were significant differences in differentiation degree and adventitious bud morphology. Combined with Table 3 and Fig. 3， it can be seen that when the cytokinin concentration was high， the morphology of adventitious buds formed by redifferentiation was abnormal. Therefore， after primary culture and induction of callus， appropriate reduction of cytokinin concentration is conducive to adventitious bud differentiation， in which 6BA and ZT concentrations should not exceed 0.5 mg/L and TDZ concentration should not exceed 0.05 mg/L.
Xiuxiu LI et al. Study on Highfrequency Callus Induction From Aseptic Plantlets of Anthuium andraeanum
Discussion
In the research and application of A. andraeanum tissue culture and rapid propagation， the plants are mainly regenerated by the organogenesis pathway of callus， and most of the explants are from potted seedlings or aseptic seedlings[8-9]. Liu[10]induced calli using the leaves， petioles and roots of the aseptic seedlings of A. andraeanum ‘Pink champion’ as explants， and the average callus production rate of the leaves was 70.1%， which was higher than those of the petioles and root segments. However， in this study， the callus production rate of the leaves from the ‘Alabama’ aseptic plantlets was only 46.7%. The reason for this difference may be related to A. andraeanum varieties， culture media and explant treatment methods. The leaf explants were free of removal of leaf opex and had a small wound area， resulting in a lower callus production rate.
The callus induction effect of A. andraeanum is also affected by its genotype and explant treatment. For example， among the test varieties ‘Robino’， ‘Pink champion’， ‘Champion’， ‘ Sweetheart Red’， ‘Arizona’ and ‘Moli’， the callus induction rate of the ‘Champion’ aseptic seedlings can reach 96.7%， while ‘Moli’ failed to induce callus[10]. Such three varieties as ‘Kratt’， ‘Arizona’ and ‘Atlanta’ all showed the callus induction rates of stem segments with petioles reaching 63% or more[11]. In this study， when the callus induction was carried out with the stem segments as the explants， the four varieties， ‘Pink champion’， ‘Turenza’， ‘Alabama’ and ‘Yalihong’ all exhibited the callus production rate of 100%， and ‘Madural ’ and ‘Hongchun’ showed the callus production rates higher than 70%， indicating that the callus induction rate had been significantly improved， which might be related to their genotypes and the explant treatment method. In this study， the excised stem explants retained the upper growth point and two young leaves， suggesting that the combination of endogenous and exogenous phytohormones would facilitate the formation of callus. This study also showed that a high concentration of thidiazepine （TDZ） can cause the malformation of the differentiated adventitious buds， which is similar to the test results of Wang et al.[12]， that is， the induction rate of A. andraeanum callus increases with the concentration of TDZ increasing， but too high concentration is teratogenic to adventitious bud differentiation. In this study， when the concentration of TDZ was 2.0mg/L in the primary culture medium， various varieties still had a better callus induction effect， but after being transferred to the medium with the TDZ concentration of 0.05 mg/L， the adventitious buds were manifested as malformed plantlets. Therefore， in the process of inducing dedifferentiation and adventitious bud redifferentiation of A. andraeanum callus， the concentration of TDZ should not be too high.
References
[1] WU AL. Tissue culture in vitro and rapid propagation of Anthurium androaeanum[J]. Genomics and Applied Biology， 2010， 29（1）： 185-190. （in Chinese）
[2] DANG CJ， ZHOU ZQ， LIU C， et al. Tissue culture and reproduction of Anthurium varieties[J]. Journal of Agricultural Sciences， 2015， （1）： 61-64. （in Chinese）
[3] FINNIE JF， STADEN JV. Invitro culture of Anthurium andraeanum[J]. South African Journal of Botany， 1986， 52（4）：343-346.
[4] NIU RH. Construction of Anthuium andraeanum genetic transformation system and antibacterial peptide gene expression vector[D]. Suzhou： Suzhou University， 2015： 13-17. （in Chinese）
[5] CAI N. Optimization of largescale invitro culture techniques for Anthuium andraeanum[D]. changsha： agricultural university of Hunan， 2004： 14-21. （in Chinese）
[6] DU JR. Study on tissue culture technology of Anthuium andraeanum[D]. baoding： agricultural university of Hebei， 2010：14-23. （in Chinese）
[7] HUANG LF. Technical optimization on factory production of Anthuium andraeanum tissue culture seedlings[D]. changsha： agricultural university of Hunan， 2008： 15-23. （in Chinese）
[8] JIA Y， MA Y， GUO Y， et al. Study on tissue culture of Anthurium andraeanum Lind[J]. Journal of Henan Normal University， 2007， 35（1）：164-166.
[9] LAN QY， LI QR， HE HY， et al. The callus induction of Anthurium andraeanum Linden and bud differentiation[J]. Acta Horticulturae Sinica， 2003， （1）：107-109. （in Chinese）
[10] LIU BJ. Agrobacteriummediated Transformation of AtCBF3 and PaFT Genes into Anthurium andraeanum[D]. Wuhan： Huazhong Agricultural University， 2011： 14-21. （in Chinese）
[11] YAO Z. Genetic transformation of Anthurium andraeanum and Phalaenopsis aphrodite Rchb. F. by carotenoid synthase genes PSY， PDS， LycB and LycE[D]. Changchun： Jilin University， 2005： 23-28. （in Chinese）
[12] WANG Y， REN M， YANG Y. Application of thidiazuron on tissue culture in Anthurium andraeanum[J]. Tianjin Agricultural Sciences， 2013， 19（9）： 12-14. （in Chinese）
Editor： Yingzhi GUANG Proofreader： Xinxiu ZHU