ÐÎÑÑÈÉÑÊÀß ÔÅÄÅÐÀÖÈß RU (19) (11) 2 279 212 (13) C2 (51) ÌÏÊ A01H 4/00 (2006.01) ÔÅÄÅÐÀËÜÍÀß ÑËÓÆÁÀ ÏÎ ÈÍÒÅËËÅÊÒÓÀËÜÍÎÉ ÑÎÁÑÒÂÅÍÍÎÑÒÈ, ÏÀÒÅÍÒÀÌ È ÒÎÂÀÐÍÛÌ ÇÍÀÊÀÌ (12) ÎÏÈÑÀÍÈÅ ÈÇÎÁÐÅÒÅÍÈß Ê ÏÀÒÅÍÒÓ (21), (22) Çà âêà: 2004121131/13, 13.07.2004 (72) Àâòîð(û): Äåìåíêî Âàñèëèé Èâàíîâè÷ (RU) (24) Äàòà íà÷àëà îòñ÷åòà ñðîêà äåéñòâè ïàòåíòà: 13.07.2004 (43) Äàòà ïóáëèêàöèè çà âêè: 10.01.2006 Àäðåñ äë ïåðåïèñêè: 127550, Ìîñêâà, óë. Òèìèð çåâñêà , 49, ÔÃÎÓ ÂÏÎ ÐÃÀÓ-ÌÑÕÀ èì. Ê.À. Òèìèð çåâà, ïàòåíòíûé îòäåë âåðõíèé - èç òåðìîóñàäî÷íîé ïëåíêè, êîòîðóþ ïîñëå óêîðåíåíè ðàñòåíèé óäàë þò. Çà ñ÷åò äàëüíåéøåé àêêëèìàòèçàöèè íàäçåìíîé ñèñòåìû ðàñòåíèé äî èõ ïåðåñàäêè â íåñòåðèëüíûå óñëîâè óâåëè÷èâàåòñ âûõîä ðàçìíîæåííûõ in vitro ðàñòåíèé. 1 òàáë. R U 2 2 7 9 2 1 2 (57) Ðåôåðàò: Èçîáðåòåíèå îòíîñèòñ ê âûðàùèâàíèþ ðàñòåíèé. Ïîñëå âûñàäêè ïîáåãîâ â ñîñóäû ñ ïèòàòåëüíîé ñðåäîé ñîñóäû çàêðûâàþò äâóì ñëî ìè ïëåíêè. Áëèæíèé ê ðàñòåíè ì ñëîé âûïîëíåí èç ïëåíêè, ïðîïóñêàþùåé âëàãó, à Ñòðàíèöà: 1 RU C 2 C 2 (54) ÑÏÎÑÎÁ ÐÀÇÌÍÎÆÅÍÈß ÐÀÑÒÅÍÈÉ in vitro 2 2 7 9 2 1 2 (56) Ñïèñîê äîêóìåíòîâ, öèòèðîâàííûõ â îò÷åòå î ïîèñêå: SU 1400557 A1, 19.12.1985. SU 1340652 A1, 30.09.1987. RU 2201071 C1, 27.03.2003. R U (45) Îïóáëèêîâàíî: 10.07.2006 Áþë. ¹ 19 (73) Ïàòåíòîîáëàäàòåëü(è): ÔÅÄÅÐÀËÜÍÎÅ ÃÎÑÓÄÀÐÑÒÂÅÍÍÎÅ ÎÁÐÀÇÎÂÀÒÅËÜÍÎÅ Ó×ÐÅÆÄÅÍÈÅ ÂÛÑØÅÃÎ ÏÐÎÔÅÑÑÈÎÍÀËÜÍÎÃÎ ÎÁÐÀÇÎÂÀÍÈß "ÐÎÑÑÈÉÑÊÈÉ ÃÎÑÓÄÀÐÑÒÂÅÍÍÛÉ ÀÃÐÀÐÍÛÉ ÓÍÈÂÅÐÑÈÒÅÒ - ÌÑÕÀ èìåíè Ê.À.ÒÈÌÈÐßÇÅÂÀ" (RU) RUSSIAN FEDERATION RU (19) (11) 2 279 212 (13) C2 (51) Int. Cl. A01H 4/00 (2006.01) FEDERAL SERVICE FOR INTELLECTUAL PROPERTY, PATENTS AND TRADEMARKS (12) ABSTRACT OF INVENTION (21), (22) Application: 2004121131/13, 13.07.2004 (72) Inventor(s): Demenko Vasilij Ivanovich (RU) (24) Effective date for property rights: 13.07.2004 (43) Application published: 10.01.2006 (54) METHOD FOR in-vitro MULTIPLICATION OF PLANTS shrinkable film. After plant rooting, heat shrinkable film is removed. EFFECT: increased yield of in-vitro multiplied plants due to further acclimatization of aboveground part of plants. 1 tbl, 1 ex R U 2 2 7 9 2 1 2 C 2 C 2 (57) Abstract: FIELD: plant growing. SUBSTANCE: method involves planting sprouts in reservoirs comprising nutritive medium and closing sprouts with two film layers. Layer closest to plants is made from moisture permeable film, and upper layer is made from heat Ñòðàíèöà: 2 EN 2 2 7 9 2 1 2 Mail address: 127550, Moskva, ul. Timirjazevskaja, 49, FGOU VPO RGAU-MSKhA im. K.A. Timirjazeva, patentnyj otdel R U (45) Date of publication: 10.07.2006 Bull. 19 (73) Proprietor(s): FEDERAL'NOE GOSUDARSTVENNOE OBRAZOVATEL'NOE UChREZhDENIE VYSShEGO PROFESSIONAL'NOGO OBRAZOVANIJa "ROSSIJSKIJ GOSUDARSTVENNYJ AGRARNYJ UNIVERSITET - MSKhA imeni K.A.TIMIRJaZEVA" (RU) RU 2 279 212 C2 5 10 15 20 25 30 35 40 Ïðåäëàãàåìîå èçîáðåòåíèå îòíîñèòñ ê îáëàñòè ñåëüñêîãî õîç éñòâà è ìîæåò áûòü èñïîëüçîâàíî â ïëîäîâûõ ïèòîìíèêàõ. Èçâåñòåí ñïîñîá ðàçìíîæåíè ðàñòåíèé in vitro (Äåìåíêî Â.È. Ìèêðîêëîíàëüíîå ðàçìíîæåíèå ïëîäîâûõ è ãîäíûõ êóëüòóð. Ì.: Èçäàòåëüñòâî ÌÑÕÀ, 1997). Îñíîâíûì íåäîñòàòêîì óêàçàííîãî ñïîñîáà âë åòñ íèçêèé ïðîöåíò ïðèæèâàåìîñòè ðàñòåíèé â íåñòåðèëüíûõ óñëîâè õ. Íàèáîëåå áëèçêèì òåõíè÷åñêèì ðåøåíèåì ê çà âë åìîìó èçîáðåòåíèþ âë åòñ ñïîñîá, ïðåäóñìàòðèâàþùèé ïîêðûòèå ñîñóäîâ ïëåíêîé Parafilm. Îäíàêî ïðè èñïîëüçîâàíèè ïëåíêè Parafilm ðàçâèâàþòñ ñâåðõîâîäíåííûå ðàñòåíè , êîòîðûå ïðè ïåðåñàäêå â íåñòåðèëüíûå óñëîâè ïîãèáàþò, âñëåäñòâèå ÷åãî ïðîöåíò ïðèæèâøèõñ ðàñòåíèé ñíèæàåòñ (Thomas M. Contolling vitrification of petunia in vitro. Programm and Abstracts 1988, p.758). Çàäà÷à ïðåäëàãàåìîãî èçîáðåòåíè - óâåëè÷èòü âûõîä ðàñòåíèé, ðàçìíîæåííûõ in vitro, çà ñ÷åò àêêëèìàòèçàöèè íàäçåìíîé ñèñòåìû åùå äî ïåðåñàäêè â íåñòåðèëüíûå óñëîâè . Äë ðåøåíè óêàçàííîé çàäà÷è ñîñóäû ñ ïîáåãàìè äë óêîðåíåíè çàêðûâàþò äâóì ñëî ìè ïëåíêè. Ïåðâûé ñëîé èç ïëåíêè, ïðîïóñêàþùåé âëàãó, íàïðèìåð öåëëîôàí, âòîðîé ñëîé - ïëåíêè, íå ïîãëîùàþùåé âëàãó, íàïðèìåð òåðìîóñàäî÷íà ïëåíêà. Ïîñëå ÷åãî ðàñòåíè îñòàâë þò íà óêîðåíåíèå. Ïîñëå óêîðåíåíè ïîáåãîâ âåðõíþþ ïëåíêó óäàë þò, ñðåäà ïîñòåïåííî âûñûõàåò, íàäçåìíà ñèñòåìà àêêëèìàòèçèðóåòñ , ãðèáíà èíôåêöè íà ïîâåðõíîñòè íå ðàçâèâàåòñ , ðàñòåíè õîðîøî ïðèæèâàþòñ â íåñòåðèëüíûõ óñëîâè õ. Ïðèìåð âûïîëíåíè ñïîñîáà. Äë ðàçìíîæåíè in vitro èñïîëüçîâàëè çåìë íèêó ñîðòà Ðåäãîíòëèò. Âåðõóøêè ïîáåãîâ çåìë íèêè âûñàæèâàëè íà ïèòàòåëüíóþ ñðåäó Ìóðàñèãà è Ñêóãà, ñîäåðæàùóþ 0,5 ìã/ë ÁÀÏ+0,01 ìã/ë ÈÌÊ+30 ã/ë ñàõàðîçû, âèòàìèíû (ïèðèäîêñèí, òèàìèí, íèêîòèíîâà êèñëîòà ïî 0,5 ìã/ë), ìåçîèíîçèò - 100 ìã/ë, àãàð 6 ã/ë. Ïîñëå ï òîãî ïàññàæà êîíãëîìåðàò ïîáåãîâ äåëèëè íà îòäåëüíûå ïîáåãè, âûñàæèâàëè íà ïèòàòåëüíóþ ñðåäó, ñîäåðæàùóþ 1/2 ìàêðîñîëåé M. Ñ., 1 ìèêðîñîëåé M. Ñ., ñàõàðîçó 20 ã/ë, 0,2 ìã/ë ÈÌÊ. Îñòàëüíûå êîìïîíåíòû, êàê íà ýòàïå ïðîëèôåðàöèè ïîáåãîâ. Óêîðåíåíèå ïðîõîäèëî ïðè 16-÷àñîâîì äíå, èíòåíñèâíîñòè ñâåòà 3-4 òûñ. ëþêñ, òåìïåðàòóðå 24-25°Ñ, ïðîäîëæèòåëüíîñòü óêîðåíåíè - 30 äíåé. ×àñòü ñîñóäîâ äë ñðàâíåíè ñ ïðåäëîæåííûì ñïîñîáîì çàêðûâàëè ïëåíêîé Parafilm. Äðóãóþ ÷àñòü çàêðûâàëè äâóì ñëî ìè ïëåíêè, ïðè÷åì ïåðâûé ñëîé ñîñòî ë èç ïëåíêè, ïðîïóñêàþùåé âëàãó (öåëëîôàíà), à âòîðîé ñëîé - èç òåðìîóñàäî÷íîé ïëåíêè, íå ïðîïóñêàþùåé âëàãó. Ïîñëå óêîðåíåíè ðàñòåíè , çàêðûòûå ïëåíêîé Parafilm, ïåðåñàæèâàëè: 1. íåïîñðåäñòâåííî â íåñòåðèëüíûå óñëîâè , ïðîöåíò ïðèæèâøèõñ ðàñòåíèé ñîñòàâèë 50%; 2. ïîñëå óäàëåíè ïëåíêè Parafilm ñîñóäû îñòàâë ëè îòêðûòûìè â òå÷åíèå 10 äíåé. Ïîñëå ïåðåñàäêè â íåñòåðèëüíûå óñëîâè ðàñòåíè ïîðàæàëà ãðèáíà èíôåêöè . Ó óêîðåíèâøèõñ ðàñòåíèé â ñîñóäàõ, çàêðûòûõ äâóì ñëî ìè ïëåíêè, ñíèìàëè âåðõíèé ñëîé èç òåðìîóñàäî÷íîé ïëåíêè è îñòàâë ëè ðàñòåíè äë àêêëèìàòèçàöèè ïîä îäíèì ñëîåì ïðîïóñêàþùåé âëàãó ïëåíêè. Ïîñëå çàâåðøåíè àêêëèìàòèçàöèè ðàñòåíè ïåðåñàæèâàëè â íåñòåðèëüíûå óñëîâè . Ïðîöåíò ïðèæèâøèõñ ðàñòåíèé ñîñòàâèë 89-90%. Òàáëèöà Âëè íèå òèïà ïîêðûòè íà ïðèæèâàåìîñòü çåìë íèêè â íåñòåðèëüíûõ óñëîâè õ 45 Òèï ïîêðûòè Óêîðåí åìîñòü, % Ïðèæèâàåìîñòü â íåñòåðèëüíûõ óñëîâè õ, % Èçâåñòíûé ñïîñîá: ïëåíêà Parafilm 95 50 Ïîñëå óêîðåíåíè ïëåíêà ñíèìàåòñ , ñîñóäû îòêðûòû 10 äíåé 95 Ãðèáíà èíôåêöè Ïðåäëàãàåìûé ñïîñîá: öåëëîôàí-òåðìîóñàäî÷íà ïëåíêà 89 95 Òàêèì îáðàçîì, ïðåäëàãàåìîå èçîáðåòåíèå ïîçâîëèò çíà÷èòåëüíî óâåëè÷èòü ïðèæèâàåìîñòü ðàñòåíèé â íåñòåðèëüíûõ óñëîâè õ. 50 Ôîðìóëà èçîáðåòåíè Ñïîñîá ðàçìíîæåíè ðàñòåíèé in vitro, âêëþ÷àþùèé óêîðåíåíèå ïîáåãîâ â ñòåðèëüíûõ óñëîâè õ â çàêðûòûõ ñîñóäàõ ñ ïèòàòåëüíîé ñðåäîé ñ ïîñëåäóþùèì âûðàùèâàíèåì Ñòðàíèöà: 3 DE CL RU 2 279 212 C2 5 ðàñòåíèé â íåñòåðèëüíûõ óñëîâè õ, îòëè÷àþùèéñ òåì, ÷òî ïîñëå âûñàäêè ïîáåãîâ â ïèòàòåëüíóþ ñðåäó ñîñóäû çàêðûâàþò äâóì ñëî ìè ïëåíêè, ïðè÷åì ïåðâûé, áëèæíèé ê ðàñòåíè ì ñëîé âûïîëíåí èç ïëåíêè, ïðîïóñêàþùåé âëàãó, à âòîðîé âåðõíèé - èç òåðìîóñàäî÷íîé ïëåíêè, êîòîðóþ ïîñëå óêîðåíåíè ðàñòåíèé óäàë þò, à ïåðåñàäêó îñóùåñòâë þò ïîñëå èõ àêêëèìàòèçàöèè ïîä ïëåíêîé, ïðîïóñêàþùåé âëàãó. 10 15 20 25 30 35 40 45 50 Ñòðàíèöà: 4