باززايي درون شيشه‌اي گياه زعفران زينتي (Crocus vernus L.) با استفاده از تنظيم‌كننده‌هاي رشد گياهي

In Vitro Regeneration of Ornamental Crocus (Crocus vernus L.) by Using Plant Growth Regulators

زﻋﻔﺮان زﯾﻨﺘﯽ از ﺗﯿﺮه زﻧﺒﻘﯿﺎن )Iridaceae( ﯾﮑﯽ از ﻣﻬﻢ ﺗﺮﯾﻦ ﮔﯿﺎﻫﺎن ﭘﯿﺎزي زﯾﻨﺘﯽ اﺳﺖ. ﺗﮑﺜﯿﺮ اﯾﻦ ﮔﯿﺎه ﺑﻪ وﺳﯿﻠﻪ ﭘﺪاژه ﺑﻪ دﻟﯿﻞ اﻧﺘﻘﺎل اﻧﻮاع ﺑﯿﻤﺎري ﻫﺎي ﻗﺎرﭼﯽ و ﺑﺎﮐﺘﺮﯾﺎﺋﯽ و ﻧﯿﺰ دوره ﺧﻔﺘﮕﯽ ﻃﻮﻻﻧﯽ 4 ﺗﺎ 5 ﻣﺎﻫﻪ، از ﻧﻈﺮ ﺗﺠﺎري ﻣﻘﺮون ﺑﻪ ﺻﺮﻓﻪ ﻧﻤﯽ ﺑﺎﺷﺪ. ﺑﻨﺎﺑﺮاﯾﻦ ﺗﮑﺜﯿﺮ زﻋﻔﺮان زﯾﻨﺘﯽ از ﻃﺮﯾﻖ ﮐﺸﺖ درون ﺷﯿﺸﻪ اي راﻫﮑﺎر ﻣﻨﺎﺳﺒﯽ ﺑﺮاي ﺗﮑﺜﯿﺮ ﺗﺠﺎري آن اﺳﺖ. ﺑﻪ ﻣﻨﻈﻮر رﯾﺰازدﯾﺎدي زﻋﻔﺮان زﯾﻨﺘﯽ، اﺛﺮ ﻣﺤﯿﻂ ﻫﺎي ﮐﺸﺖ ﻣﺨﺘﻠﻒ در آزﻣﺎﯾﺶ اﻧﺪام زاﯾﯽ ﻣﺴﺘﻘﯿﻢ ﺑﻪ ﺻﻮرت ﻓﺎﮐﺘﻮرﯾﻞ در ﻗﺎﻟﺐ ﻃﺮح ﮐﺎﻣﻼً ﺗﺼﺎدﻓﯽ در 3 ﺗﮑﺮار در آزﻣﺎﯾﺸﮕﺎه ﮐﺸﺖ ﺑﺎﻓﺖ ﮔﺮوه ﻋﻠﻮم ﺑﺎﻏﺒﺎﻧﯽ داﻧﺸﮕﺎه ﻋﻠﻮم ﮐﺸﺎورزي و ﻣﻨﺎﺑﻊ ﻃﺒﯿﻌﯽ ﺧﻮزﺳﺘﺎن، در ﺳﺎل 1394-95 اﻧﺠﺎم ﺷﺪ. رﯾﺰﻧﻤﻮﻧﻪ ﻫﺎي ﻣﻮرد اﺳﺘﻔﺎده در اﯾﻦ آزﻣﺎﯾﺶ ﺟﻮاﻧﻪ ﻫﺎي اﻧﺘﻬﺎﯾﯽ و ﺟﺎﻧﺒﯽ ﺑﻮدﻧﺪ ﮐﻪ در 10 ﺗﯿﻤﺎر ﻫﻮرﻣﻮﻧﯽ و ﮐﻨﺘﺮل در ﻣﺤﯿﻂ ﮐﺸﺖ MS ﻣﻮرد آزﻣﺎﯾﺶ ﻗﺮار ﮔﺮﻓﺘﻨﺪ. ﺑﺮاي ﮔﻨﺪزداﯾﯽ ﺳﻄﺤﯽ رﯾﺰﻧﻤﻮﻧﻪ ﻫﺎ ﭘﺲ از ﻗﺮار ﮔﯿﺮي در زﯾﺮ آب ﺟﺎري ﺑﻪ ﻣﺪت 30 دﻗﯿﻘﻪ از ﺗﯿﻤﺎر ﻗﺎرچ ﮐﺶ ﺑﻨﻠﯿﺖ ﻣﺤﻠﻮل در آب ﮔﺮم ﺑﻪ ﻣﺪت 30 دﻗﯿﻘﻪ، اﺗﺎﻧﻮل 70 درﺻﺪ ﺑﻪ ﻣﺪت 30 ﺛﺎﻧﯿﻪ و ﺳﻪ ﺑﺎر آﺑﮑﺸﯽ در آب ﻣﻘﻄﺮ اﺳﺘﺮﯾﻞ و ﻫﯿﭙﻮﮐﻠﺮﯾﺪ ﺳﺪﯾﻢ 15 درﺻﺪ ﺑﻪ ﻣﺪت 6 دﻗﯿﻘﻪ و ﺳﻪ ﺑﺎر آب ﮐﺸﯽ اﺳﺘﻔﺎده ﺷﺪ. ﺗﯿﻤﺎرﻫﺎي ﻫﻮرﻣﻮﻧﯽ ﻣﻮرد اﺳﺘﻔﺎده ﺷﺎﻣﻞ BAP )1 ،0/5 و 2 ﻣﯿﻠﯽ ﮔﺮم در ﻟﯿﺘﺮ(، TDZ )0/5، 0/075 و 1 ﻣﯿﻠﯽ ﮔﺮم در ﻟﯿﺘﺮ( و Kin )1 ،0/5 و 2 ﻣﯿﻠﯽ ﮔﺮم در ﻟﯿﺘﺮ( ﻫﻤﺮاه ﺑﺎ 0/1 ﻣﯿﻠﯽ ﮔﺮم در ﻟﯿﺘﺮ IBA ﺑﻮدﻧﺪ . ﻧﺘﺎﯾﺞ ﻧﺸﺎن داد ﺑﯿﺸﺘﺮﯾﻦ ﻃﻮل ﺷﺎﺧﺴﺎره و ﺗﻌﺪاد آن ﺣﺎﺻﻞ از رﯾﺰﻧﻤﻮﻧﻪ ﺟﻮاﻧﻪ اﻧﺘﻬﺎﺋﯽ و ﻣﺤﯿﻂ ﮐﺸﺖ MS ﻫﻤﺮاه ﺑﺎ 0/5 ﻣﯿﻠﯽ ﮔﺮم در ﻟﯿﺘﺮ TDZ، 0/5 ﻣﯿﻠﯽ ﮔﺮم در ﻟﯿﺘﺮ BAP، 0/5 ﻣﯿﻠﯽ ﮔﺮم در ﻟﯿﺘﺮ Kin و 0/1 ﻣﯿﻠﯽ ﮔﺮم در ﻟﯿﺘﺮ IBA ﺗﻮﻟﯿﺪ ﮔﺮدﯾﺪ ﮐﻪ ﺑﺎ ﺳﺎﯾﺮ ﺗﯿﻤﺎرﻫﺎ در ﺳﻄﺢ اﺣﺘﻤﺎل 1 درﺻﺪ آزﻣﻮن داﻧﮑﻦ اﺧﺘﻼف ﻣﻌﻨﯽ دار داﺷﺖ. ﻫﻤﭽﻨﯿﻦ ﻧﺘﺎﯾﺞ ﻧﺸﺎن داد ﻣﺤﯿﻂ ﮐﺸﺖ MS ﻫﻤﺮاه ﺑﺎ 2 ﻣﯿﻠﯽ ﮔﺮم در ﻟﯿﺘﺮ BAP ﻫﻤﺮاه ﺑﺎ 0/1 ﯿﻠﯽ ﮔﺮم در ﻟﯿﺘﺮ IBA ﺑﯿﺸﺘﺮﯾﻦ اﺛﺮ را روي ﺗﻌﺪاد ﭘﺪاژك ﻫﺎي ﺗﻮﻟﯿﺪ ﺷﺪه زﻋﻔﺮان زﯾﻨﺘﯽ داﺷﺖ

Crocus flower, which belongs to Iridaceae family, is one of the most important ornamental bulbous plants. The propagation of Crocus vernus L. by corm is not commercially affordable due to transmission of fungal and bacterial diseases as well as the long dormancy period which takes 4 to 5 months. Thus, in vitro culture of Crocus vernus is a suitable solution for commercial reproduction. Plant tissue culture technique would be a better alternative for improving quality and production. Materials and Methods In order to micropropagate ornamental saffron, the effect of different culture media on direct organogenesis experiment was conducted in factorial arrangement in a completely randomized design with 3 replications in the tissue culture lab of the Department of Horticulture at Agricultural Sciences and Natural Resources University of Khuzestan from 2015 to 2016. The explants used in this experiment were latheral and terminal buds which were evaluated in 10 hormons treatments and control in the Murashige and Skoog medium (MS). To eliminate surface contamination, the explants were first immersed under water for 30 minutes after placing the corms. Next, they were placed in high temperature Benlate fungicide solution (55° C) for 30 minutes. Then, they were put in 70% ethanol (ethyl alcohol) for 30 seconds and 15% Sodium hypochlorite solution for 6 minutes. The explants were finally washed three times with sterile distilled water under the air flow bench. The effects of plant growth regulators including BAP (0.5, 1.0, 2.0 mg/l), TDZ (0.5, 0.75, 1.0 mg/l), Kin (0.5, 1.0, 2.0 mg/l) and 0.1mg/l IBA in direct regeneration were investigated. Results The results showed that the kind of plant growth regulators (PGR) and the explant type are very important in regeneration of Crocus vernus so that we can not secure any shoots without PGR. In direct organogenesis experiment, the maximum number of multiple shoots (15.84) was obtained from apical bud explant in MS medium supplemented with 5/0 mg/l TDZ along with 5/0 mg/l BAP, 5/0 mg/l KIN and 1/0 mg/l IBA, after 10 weeks, that had a significant difference with other treatments at 1% Duncan test. We observed 2.o mg/l BAP along with 1/0 mg/l IBA had the greatest effect on the number of corms. Discussion The results showed the shoot regeneration is controlled by the ratio of cytokinin with auxin. It should be mentioned that cytokinin plays a role in the synthesis of RNA, stimulation of the production of protein, and the activity of some enzymes. The results also indicated that the use of cytokinin along with auxin has a beneficial effect on shoot regeneration.