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Abstract Five different parts of Rosa damascene Mill. twigs were treated with hormone solution （50 mg/L NAA+100 mg/L IBA） and planted in base material. After 50 d of growth， the survival rate and rooting condition were investigated. The results showed that the survival rate from 5 different parts of cutting stem decreased from top to bottom （upper 98.22%， middle and upper 95.12%， middle 91.10%， middle and lower 72.90%， lower 71.12%）. The same results were also observed in the number， total length and fresh weight of secondary roots of cutting segments from different parts of twigs.
Key words Rosa damascene Mill.； Shoots； Segments； Cuttings； Survival ratio
Rosa damascene Mill. belongs to Rosa L. in Rosaceae family. It is a kind of bush， native to Syria[1]. R. damascene Mill. is the main Rosa variety cultivated in Bulgaria as "the kingdom of rose". R. damascene Mill. essential oil is an international popular odor type， honored as "liquid gold"[2]. In recent years， R. damascene Mill. is introduced in China and cultivated in various areas[3-4]， and becomes one of the crops for structural adjustment of agriculture in mountainous and hilly areas. The propagation of R. damascene Mill. is mainly performed by root division， layering， cutting and tissue culture[5]. Among the various methods， root division and layering are restricted by factors including season， labor cost and number of female parent and have the disadvantages of long time and not high efficiency， tissue culture is rapid， but has high cost， while cutting has a high efficiency and low cost， and is thus a main means for largescale propagation[6]. As to the cutting propagation of R. damascene Mill.， there have been studies about the effects of plant growth regulator and substrate on its rooting rate[7]. In this study， through the comparison of the survival rates and rooting and growth conditions of cuttings from different parts of the same twig of R. damascene Mill.， the effects of different segments of twigs as cuttings on the efficiency of cutting propagation were compared， so as to guide largescale rapid propagation of R. damascene Mill.
Materials and Methods
Cutting treatment
The experiment was carried out in the rose planting base of Xiangyang Academy of Agricultural Sciences. From twoyearold R. damascene Mill. plants， strong new semilignified twigs free of diseases and pests were selected as experimental materials. Each twig was cut into five segments from the top down as cuttings， i.e.， I： the upper part， II： the middleupper part， III： the middle part， IV： the lowermiddle part， and V： the lower part. Each segment was one treatment， and each of the five treatments had three replicates， each of which included 45 cuttings. The twigs of R. damascene Mill. were collected at 8：00 a.m. on October 7， 2016， with a length of about 60 cm. The collected twigs were sprayed with water and kept in shade， and cut into segments timely. At first， 2-3 leaves at the top of each twig were removed， and then five segments were cut off from the top down. Each segment had a length of 10 cm， and was remained with one compound leaf having 4-6 simple leaves. The cutting was started about 5 mm above the compound leaf flatly， and the bottom of each segment was cut into a slant of 45°. The cuttings were soaked into the 800 times mixed diluent of agricultural streptomycin and hymexazol， and fished up 10 min later. A suspension， 50 mg/L NAA+100 mg/L IBA， was prepared， and used for soaking the cuttings for 3 min. The cuttings were fished up and airdried for 2-3 min， and cutting was then performed timely. Cutting seedbed and substrate
At first， plastic aperture disks （51 cm× 29 cm） were numbered， each having 45 （9×5） apertures. Each number was corresponding to one replicate of cutting treatment， and according to the experiment design， the disks were randomly placed on bamboo shelves. Perlite and coco coir were mixed at a ratio of 1∶4 as substrate. The substrate was filled in nonwoven fabric bags with a bottom width of 7 cm and a height of 9 cm. One nutrition bag filled with the substrate was added into one aperture.
Cutting method and management
A small hole with depth of 3 cm was formed with a small stick in the substrate in each nutrition bag， to facilitate the insertion of cuttings. The insertion depth of cuttings was not deeper than 2/3 of the nutrition bag. After insertion， the substrate was pushed lightly， so that the cuttings closely contacted the substrate. After all the cutting was finished on the very day， the cutting seedbed was sterilized with 800 times carbendazol diluent. On the second day， the timing spraying equipment was initiated， to ensure the leaves of the cuttings wet. As to daily management， in the glasshouse， the temperature was kept in the range of 18-25 ℃， and the relative air humidity was controlled within 65%-85%.
Investigation content and data processing
After the cuttings grew for 50 d， the survival rate of cuttings was investigated， and the calculation was performed according to the formula： Survival rate=Number of rooted cuttings/Number of cuttings×100%.
From each treatment， 10 rooted plants were taken randomly， for the investigation of average root number per plant， total root length， fresh weight of root， new shoot length and number of new compound eaves. The experimental data were processed in Microsoft Office Excel 2007， and used for plotting and tabulation. Statistical analysis was performed in DPS v3.01 by Duncans new method.
Results and Analysis
Survival condition of twig segmentation cutting
The survival condition of semilignified R. damascene Mill. twigs was shown in Table 1 and Fig. 1. It could be seen from Table 1 and Fig. 1 that the survival rate of twig segmentation cutting of R. damascene Mill. showed a remarkable decreasing trend from top to bottom. The survival rates of the upper and middleupper parts were the highest， of 98.22% and 95.12% ， respectively； the middle part had a survival rate of 91.10%； and the lowermiddle part and the lower part exhibited the lowest survival rates， of 72.90% and 71.12%， respectively. The survival rates of segments of the upper and middleupper parts were significantly higher than those of the segments of other parts （P<0.05）， and the survival rate of the segments of the middle part was significantly different from those of the segments of the lowermiddle and lower parts （P<0.05）. The growth condition of the underground part of plants survived semilignified twig segmentation cutting is shown in Table 1 and Fig. 2. It could be seen from Table 1 and Fig. 2 that the average rooting number， total root length and fresh weight of roots of individual plants obtained by semilignified twig segmentation cutting of R. damascene Mill. decreased from the top down overall. The average root number of individual survived twig segmentation cutting of R. damascene Mill. was the highest in the upper part treatment， which had 48.0 roots； the middleupper and middle part treatments had the second most roots， i.e.， 42.1 and 43.1 roots， respectively； and the lowermiddle and lower part treatments had fewer roots， i.e.， 37.2 and 27.4 roots， respectively. There was a significant difference between the upper part segment and lower part segment treatments （P<0.05）. The average total root lengths of individual plants from the various segments were 107.64， 90.32， 79.18， 46.11 and 41.37 cm from the top down. The total root length of upper pat treatment was significantly different those of the middle， lowermiddle and lower part treatments （P<0.05）， and the total root lengths of the uppermiddle and middle part treatments were significantly different from those of the lowermiddle and lower part treatment （P<0.05）. The upper part treatment had the highest average fresh weight of roots， of 0.59 g； the middleupper， middle and lowermiddle part exhibited the second highest values， of 0.50， 0.51 and 0.49 g， respectively； and the lower part had the lowest fresh weight of roots， of 0.30 g. The upper， middleupper， middle and lowermiddle parts were significantly different from the lowest part in the fresh weight of roots （P<0.05）.
The growth condition of the aboveground part of plants survived semilignified twig segmentation cutting is shown in Table 1 and Fig. 3. It could be seen from Table 1 and Fig. 3 that the length of the new shoot of individual plants survived twig segmentation cutting was 6.45， 7.85， 8.60， 8.10 and 7.70 cm in the upper part， middleupper part， middle part， lowermiddle part and lower part treatments， respectively. As to the number of new compound leaves， the upper part treatment had the most leaves， 5.5 leaves， and other treatments had 4.2， 4.9， 4.6 and 4.8 leaves from top to bottom， respectively. Overall， except that the new shoot length of the middle part treatment was significantly shorter than that of the upper part treatment （P<0.05）， there were no significant differences in the growth condition between the segments of other parts （P>0.05）. Liyun LI et al. Propagation Experiment of Twig Segmentation Cutting of Rosa damascene Mill.
Conclusions and Discussion
Segments from different parts of R. damascene Mill. twigs serving as the cuttings significantly affected the efficiency of cutting propagation. The survival rate of cuttings was the highest in the upper part treatment， decreased slightly in the middleupper and middle part treatments， and was lower in the lowermiddle and lower part treatments. Comprehensively， taking the data including root number of survived plants into consideration， selecting the segments from the parts above the middle part of R. damascene Mill. twigs could greatly improve the cutting propagation efficiency of R. damascene Mill. The selection method of cuttings： On the parent plants of R. damascene Mill.， semilignified twigs newly growing out in the very year are selected， and the 30-35 cm upper part of each twig is cut into 2-3 segments as cuttings， each having a length of about 10 cm. If the cutting propagation technique is mature， the survival rate of these cuttings could reach 90%. R. chinensis Jacq. and R. damascene Mill. both belong to Rosaceae family. Li[8]showed through the experiment of different cutting parts of different Chinese rose varieties that， under the same cutting condition， the main factor affecting cutting of Chinese rose is the part of cuttings. Ma et al.[9]found through their study on raising R. rugosa Thunb. seedlings by cutting that， in the same period， the upper part of twigs had better rooting effects of cuttings than the middle and lower part segments， which accords with the results in this study.
However， the new shoot length of the upper part segments was significantly lower than the new shoot length of the middle part segments， which might be due to that the axillary buds of middle part of twigs developed early and were stronger， while the axillary buds of the upper part of the twigs developed late and were tender， the nutrition basis of the upper part segments was poorer than that of the middle part， and as a result， after the survival of the cuttings， the new shoot length of the upper part segments was shorter than the middle part segments.
References
[1]XU Y， SU C. Introduction and cultivation and industrialization prospect of Rosa damascene tyigintipetala in Panxi Area[J]. Journal of Mountain Science， 2006， 24（5）： 636-640.
[2]ZHANG R， WEI AZ. Study on Rosa damascena Mill. essential oil produced in Shangzhou[J]. Acta Botanica BorealiOccidentalia Sinica， 2005， 25（7）： 1477-1479. [3]WANG Y， TANG YH， WU DQ， et al. Introduction and cultivation of Rosa damascena Mill. var. kazanlika Anji， Zhejiang[J]. Journal of Shanghai Jiaotong University ： Agricultural Science， 2009， 27（3）： 226-230.
[4]YANG FQ. Propagation technique of Rosa damascena Mill. in Yili region[J]. Rurai Science & Thchnology， 2013（12）： 47-48.
[5]LI WY， WANG WZ. Preliminary study of rose resources in China[J]. Acta Horticulturae Sinica， 1983（3）： 213-215.
[6]XU LJ， LI ZB， JIANG SL， et al. Study on Rosa damascena Mill. tissue culture[J]. The Journal of Hebei Forestry Science and Technology， 2015（2）： 19-21.
[7]YANG LN， REN JA， WANG Y， et al. Study on rooting rate of different varieties of Rosa damascena Mill. cutting[J]. Journal of Shanxi Agricultural University： Natural Science Edition， 2012， 32（1）： 18-21.
[8]LI CH. Cutting propagation experiment of different Chinese rose varieties using different cutting parts[J]. Modern Horticulture， 2015（3）： 18-19.
[9]MA C， YUAN F. Effects of cutting collection period and part on quality of raising seedlings by cutting of Chinese rose[J]. Horticulture & Seed， 2011（3）： 17-18.
Editor： Yingzhi GUANG Proofreader： Xinxiu ZHU
Key words Rosa damascene Mill.； Shoots； Segments； Cuttings； Survival ratio
Rosa damascene Mill. belongs to Rosa L. in Rosaceae family. It is a kind of bush， native to Syria[1]. R. damascene Mill. is the main Rosa variety cultivated in Bulgaria as "the kingdom of rose". R. damascene Mill. essential oil is an international popular odor type， honored as "liquid gold"[2]. In recent years， R. damascene Mill. is introduced in China and cultivated in various areas[3-4]， and becomes one of the crops for structural adjustment of agriculture in mountainous and hilly areas. The propagation of R. damascene Mill. is mainly performed by root division， layering， cutting and tissue culture[5]. Among the various methods， root division and layering are restricted by factors including season， labor cost and number of female parent and have the disadvantages of long time and not high efficiency， tissue culture is rapid， but has high cost， while cutting has a high efficiency and low cost， and is thus a main means for largescale propagation[6]. As to the cutting propagation of R. damascene Mill.， there have been studies about the effects of plant growth regulator and substrate on its rooting rate[7]. In this study， through the comparison of the survival rates and rooting and growth conditions of cuttings from different parts of the same twig of R. damascene Mill.， the effects of different segments of twigs as cuttings on the efficiency of cutting propagation were compared， so as to guide largescale rapid propagation of R. damascene Mill.
Materials and Methods
Cutting treatment
The experiment was carried out in the rose planting base of Xiangyang Academy of Agricultural Sciences. From twoyearold R. damascene Mill. plants， strong new semilignified twigs free of diseases and pests were selected as experimental materials. Each twig was cut into five segments from the top down as cuttings， i.e.， I： the upper part， II： the middleupper part， III： the middle part， IV： the lowermiddle part， and V： the lower part. Each segment was one treatment， and each of the five treatments had three replicates， each of which included 45 cuttings. The twigs of R. damascene Mill. were collected at 8：00 a.m. on October 7， 2016， with a length of about 60 cm. The collected twigs were sprayed with water and kept in shade， and cut into segments timely. At first， 2-3 leaves at the top of each twig were removed， and then five segments were cut off from the top down. Each segment had a length of 10 cm， and was remained with one compound leaf having 4-6 simple leaves. The cutting was started about 5 mm above the compound leaf flatly， and the bottom of each segment was cut into a slant of 45°. The cuttings were soaked into the 800 times mixed diluent of agricultural streptomycin and hymexazol， and fished up 10 min later. A suspension， 50 mg/L NAA+100 mg/L IBA， was prepared， and used for soaking the cuttings for 3 min. The cuttings were fished up and airdried for 2-3 min， and cutting was then performed timely. Cutting seedbed and substrate
At first， plastic aperture disks （51 cm× 29 cm） were numbered， each having 45 （9×5） apertures. Each number was corresponding to one replicate of cutting treatment， and according to the experiment design， the disks were randomly placed on bamboo shelves. Perlite and coco coir were mixed at a ratio of 1∶4 as substrate. The substrate was filled in nonwoven fabric bags with a bottom width of 7 cm and a height of 9 cm. One nutrition bag filled with the substrate was added into one aperture.
Cutting method and management
A small hole with depth of 3 cm was formed with a small stick in the substrate in each nutrition bag， to facilitate the insertion of cuttings. The insertion depth of cuttings was not deeper than 2/3 of the nutrition bag. After insertion， the substrate was pushed lightly， so that the cuttings closely contacted the substrate. After all the cutting was finished on the very day， the cutting seedbed was sterilized with 800 times carbendazol diluent. On the second day， the timing spraying equipment was initiated， to ensure the leaves of the cuttings wet. As to daily management， in the glasshouse， the temperature was kept in the range of 18-25 ℃， and the relative air humidity was controlled within 65%-85%.
Investigation content and data processing
After the cuttings grew for 50 d， the survival rate of cuttings was investigated， and the calculation was performed according to the formula： Survival rate=Number of rooted cuttings/Number of cuttings×100%.
From each treatment， 10 rooted plants were taken randomly， for the investigation of average root number per plant， total root length， fresh weight of root， new shoot length and number of new compound eaves. The experimental data were processed in Microsoft Office Excel 2007， and used for plotting and tabulation. Statistical analysis was performed in DPS v3.01 by Duncans new method.
Results and Analysis
Survival condition of twig segmentation cutting
The survival condition of semilignified R. damascene Mill. twigs was shown in Table 1 and Fig. 1. It could be seen from Table 1 and Fig. 1 that the survival rate of twig segmentation cutting of R. damascene Mill. showed a remarkable decreasing trend from top to bottom. The survival rates of the upper and middleupper parts were the highest， of 98.22% and 95.12% ， respectively； the middle part had a survival rate of 91.10%； and the lowermiddle part and the lower part exhibited the lowest survival rates， of 72.90% and 71.12%， respectively. The survival rates of segments of the upper and middleupper parts were significantly higher than those of the segments of other parts （P<0.05）， and the survival rate of the segments of the middle part was significantly different from those of the segments of the lowermiddle and lower parts （P<0.05）. The growth condition of the underground part of plants survived semilignified twig segmentation cutting is shown in Table 1 and Fig. 2. It could be seen from Table 1 and Fig. 2 that the average rooting number， total root length and fresh weight of roots of individual plants obtained by semilignified twig segmentation cutting of R. damascene Mill. decreased from the top down overall. The average root number of individual survived twig segmentation cutting of R. damascene Mill. was the highest in the upper part treatment， which had 48.0 roots； the middleupper and middle part treatments had the second most roots， i.e.， 42.1 and 43.1 roots， respectively； and the lowermiddle and lower part treatments had fewer roots， i.e.， 37.2 and 27.4 roots， respectively. There was a significant difference between the upper part segment and lower part segment treatments （P<0.05）. The average total root lengths of individual plants from the various segments were 107.64， 90.32， 79.18， 46.11 and 41.37 cm from the top down. The total root length of upper pat treatment was significantly different those of the middle， lowermiddle and lower part treatments （P<0.05）， and the total root lengths of the uppermiddle and middle part treatments were significantly different from those of the lowermiddle and lower part treatment （P<0.05）. The upper part treatment had the highest average fresh weight of roots， of 0.59 g； the middleupper， middle and lowermiddle part exhibited the second highest values， of 0.50， 0.51 and 0.49 g， respectively； and the lower part had the lowest fresh weight of roots， of 0.30 g. The upper， middleupper， middle and lowermiddle parts were significantly different from the lowest part in the fresh weight of roots （P<0.05）.
The growth condition of the aboveground part of plants survived semilignified twig segmentation cutting is shown in Table 1 and Fig. 3. It could be seen from Table 1 and Fig. 3 that the length of the new shoot of individual plants survived twig segmentation cutting was 6.45， 7.85， 8.60， 8.10 and 7.70 cm in the upper part， middleupper part， middle part， lowermiddle part and lower part treatments， respectively. As to the number of new compound leaves， the upper part treatment had the most leaves， 5.5 leaves， and other treatments had 4.2， 4.9， 4.6 and 4.8 leaves from top to bottom， respectively. Overall， except that the new shoot length of the middle part treatment was significantly shorter than that of the upper part treatment （P<0.05）， there were no significant differences in the growth condition between the segments of other parts （P>0.05）. Liyun LI et al. Propagation Experiment of Twig Segmentation Cutting of Rosa damascene Mill.
Conclusions and Discussion
Segments from different parts of R. damascene Mill. twigs serving as the cuttings significantly affected the efficiency of cutting propagation. The survival rate of cuttings was the highest in the upper part treatment， decreased slightly in the middleupper and middle part treatments， and was lower in the lowermiddle and lower part treatments. Comprehensively， taking the data including root number of survived plants into consideration， selecting the segments from the parts above the middle part of R. damascene Mill. twigs could greatly improve the cutting propagation efficiency of R. damascene Mill. The selection method of cuttings： On the parent plants of R. damascene Mill.， semilignified twigs newly growing out in the very year are selected， and the 30-35 cm upper part of each twig is cut into 2-3 segments as cuttings， each having a length of about 10 cm. If the cutting propagation technique is mature， the survival rate of these cuttings could reach 90%. R. chinensis Jacq. and R. damascene Mill. both belong to Rosaceae family. Li[8]showed through the experiment of different cutting parts of different Chinese rose varieties that， under the same cutting condition， the main factor affecting cutting of Chinese rose is the part of cuttings. Ma et al.[9]found through their study on raising R. rugosa Thunb. seedlings by cutting that， in the same period， the upper part of twigs had better rooting effects of cuttings than the middle and lower part segments， which accords with the results in this study.
However， the new shoot length of the upper part segments was significantly lower than the new shoot length of the middle part segments， which might be due to that the axillary buds of middle part of twigs developed early and were stronger， while the axillary buds of the upper part of the twigs developed late and were tender， the nutrition basis of the upper part segments was poorer than that of the middle part， and as a result， after the survival of the cuttings， the new shoot length of the upper part segments was shorter than the middle part segments.
References
[1]XU Y， SU C. Introduction and cultivation and industrialization prospect of Rosa damascene tyigintipetala in Panxi Area[J]. Journal of Mountain Science， 2006， 24（5）： 636-640.
[2]ZHANG R， WEI AZ. Study on Rosa damascena Mill. essential oil produced in Shangzhou[J]. Acta Botanica BorealiOccidentalia Sinica， 2005， 25（7）： 1477-1479. [3]WANG Y， TANG YH， WU DQ， et al. Introduction and cultivation of Rosa damascena Mill. var. kazanlika Anji， Zhejiang[J]. Journal of Shanghai Jiaotong University ： Agricultural Science， 2009， 27（3）： 226-230.
[4]YANG FQ. Propagation technique of Rosa damascena Mill. in Yili region[J]. Rurai Science & Thchnology， 2013（12）： 47-48.
[5]LI WY， WANG WZ. Preliminary study of rose resources in China[J]. Acta Horticulturae Sinica， 1983（3）： 213-215.
[6]XU LJ， LI ZB， JIANG SL， et al. Study on Rosa damascena Mill. tissue culture[J]. The Journal of Hebei Forestry Science and Technology， 2015（2）： 19-21.
[7]YANG LN， REN JA， WANG Y， et al. Study on rooting rate of different varieties of Rosa damascena Mill. cutting[J]. Journal of Shanxi Agricultural University： Natural Science Edition， 2012， 32（1）： 18-21.
[8]LI CH. Cutting propagation experiment of different Chinese rose varieties using different cutting parts[J]. Modern Horticulture， 2015（3）： 18-19.
[9]MA C， YUAN F. Effects of cutting collection period and part on quality of raising seedlings by cutting of Chinese rose[J]. Horticulture & Seed， 2011（3）： 17-18.
Editor： Yingzhi GUANG Proofreader： Xinxiu ZHU