CN112021165A - A hybridization method of Curcuma plant - Google Patents
A hybridization method of Curcuma plant Download PDFInfo
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- CN112021165A CN112021165A CN202010705231.8A CN202010705231A CN112021165A CN 112021165 A CN112021165 A CN 112021165A CN 202010705231 A CN202010705231 A CN 202010705231A CN 112021165 A CN112021165 A CN 112021165A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G5/00—Fertilisers characterised by their form
- C05G5/20—Liquid fertilisers
Abstract
The invention discloses a hybridization method of curcuma plants. It includes hydroponic flowering, emasculation and stigma treatment and pollen treatment and pollination. The invention initiates the water culture hybridization method of the Zingiberaceae, saves land resources and improves the space utilization rate; solves the problem that the hybridization time is limited due to inconsistent flowering phases or short flowering phase overlapping time of different species of curcuma; the prepared improved Hoagland nutrient solution A can shorten the fruit development period and improve the maturing rate, the improved Hoagland nutrient solution B can prolong the single-flower pollination period and the inflorescence pollination period, and the pollen solution can improve the seed quantity. The invention provides a technical basis for promoting curcuma species crossbreeding work, provides reference significance for developing water culture crossbreeding technologies of other genera in the zingiberaceae, and also provides reference significance for developing water culture crossbreeding technologies of other bulbous plants.
Description
The technical field is as follows:
the invention belongs to the field of plant crossbreeding, and particularly relates to a method for crossbreeding curcuma plants.
Background art:
curcuma (Curcuma lin.) is a perennial herb of the family zingiberaceae, about 70 species worldwide, with more than 10 species in China in tropical regions of asia of predominant origin. Curcuma is a specific germplasm resource treasury integrating medicinal, ornamental, perfume, dye, seasoning, cosmetics, bactericide, chemical reagent and other uses, and has wide application (wudefo, 1981; liangcheng, 2019). The Chinese medicinal materials of curcuma root, zedoary and curcuma root are the rhizomes and root tubers of 5 kinds of curcuma plants of curcuma, curcuma wenyujin, curcuma longa, zedoary and curcuma kwangsiensis, etc. According to traditional Chinese medicine, the plants have the effects of resolving depression, promoting qi circulation, relieving pain, removing blood stasis, benefiting gallbladder, clearing away heart-fire, removing food retention, dredging channels and the like, and researches show that the plants have the activities of resisting cancer, resisting early pregnancy, resisting coagulation, resisting oxidation, protecting liver and the like (ZhouXin et al, 2004). The curcuma species is one of the most ornamental species in the zingiberaceae family, can be classified as bulbous flowers in flower classification due to developed rhizome, is a good material for arranging potted plants and gardens, has long and hard inflorescence stems and compact and gorgeous inflorescence, and is also suitable for being used as fresh cut flower materials (great sons and sons, et al, 2003; gaojiang clouds, et al, 2006). In addition, curcuma species are widely used as important seasonings, vegetables, flavors, natural pigments, etc., and are closely related to our lives.
The breeding work of the Curcuma species is promoted by the market demand for a large amount of Curcuma resources, and new species obtained by systematic breeding include Curcuma aromatica (Curcuma kwangsiensis cv. 'Pink') (great song monarch and the like, 2003), Curcuma agave (Curcuma kwangsiensis 'Manao') (Shengaiwu and the like, 2012), Curcuma aromatica (Curcuma kwangsiensis var. nanlingensis 'Xiangning') (Shengwuwu wuweining and the like, Curcuma aromatica (Curcuma peiolate 'Ziyu') (Xiao-Chung and the like, 2014), Curcuma erythrostica (Curcuma nankshanensis 'Hongyun') and the like. At present, interspecific hybridization of curcuma is reported, and the development of hybridization technical research can provide technical support for enriching germplasm resources of curcuma, improving variation frequency and directionally cultivating new species which have high economic value and meet market requirements.
The curcuma belongs to an insect-borne plant, the flower structure is special, pollination is not easy under natural conditions, and the natural fruiting rate is low. Artificial hybridization is an important way for promoting the breeding work of curcuma, the traditional field hybridization is limited by natural florescence, the problems of inconsistent florescence or short florescence overlapping time exist in different species, the breeding efficiency is seriously limited, the field hybridization is influenced by climate and environment, the breeding period is long, and the breeding efficiency is also limited. The artificial climate chamber is utilized to carry out hybridization work, and the interference and the limitation caused by the change of the field climate and the season can be overcome by regulating and controlling the environmental conditions. The artificial climate chamber hybridization can be used for cultivating breeding plants by using soil or a matrix, and can also be used by a water culture hybridization method, the water culture hybridization does not need to use soil or a culture matrix, the management cost of matrix preparation, disinfection, plowing, improvement and the like is saved, and the labor intensity is reduced. The cultivation of the cut and harvested field barley ears in Gem GuZhong (1979) in greenhouse water culture and hybridization, and Wang Qing bin et al (1997) reported the cultivation and hybridization of cut flower branches of poplar. The inventor utilizes the water culture cut flower branch hybridization of curcuma to cause the phenomenon of no fruit setting or no enlargement of young fruit, and can not obtain hybrid seeds, thereby indicating that the water culture hybridization of the cut flower branch is not suitable for the curcuma hybridization. The development of an efficient curcuma species hybridization method is an urgent problem to be solved for promoting curcuma species breeding, breeding new curcuma species and promoting the development of curcuma species industry.
At present, no ginger hybridization technology patent application is available.
Gaojiang cloud, Xia Yongmei, Huangjiayuan, Li Qingjun 2006, flowers of Zingiberaceae China, Beijing, science publishing house.
Guo Zhonghen 1979. hydroponics, plant journal, 2:47.
Beam Heng, Deng Jia Bin, Tongshan, Zhang Zhonghao, Daoruiwu 2019, systematic relation research progress of Curcuma plants in China, molecular plant breeding 11: 3695-.
Shengaiwu, Liu Mian, Zhang Shijun, Ye Zhu, Yi Cai Xia, hu Xiu, lan Xia.2012, the new variety of Guangxi E.C.' Agate Gui E.C. gardening academic newspaper, 39(7): 1419-.
Shengaiwu, Liu Mian, Zhang Shijun, Ye Zhu, Yi Cai Xia, Huxiu, Fa Yan Nu 2012, new species of ornamental flowers, Xiang Cheng nan Ling E Zhu, gardening bulletin, 39(11): 2335-.
Wang Qing bin, Wu Xinmin, Wei, Zhang Xiaosen, Jiang Yanlong. 1997, discussion of poplar hydroponic hybridization breeding technology, proceedings of the university academy of peony Jiang 1:9-10.
Wu Deo.1981, Curcuma of Zingiberaceae, Chinese plant journal (sixteenth volume, second volume), Beijing, scientific Press.
Great song monarch, handsome, liu monilia, fan hamming, square rigor 2003. ginger order flowers, Beijing, China forestry publishing house.
Great dawn, Liu Wen, great Song Jun, Deng Mei hong, Lin Wen hong.2014, breeding and application of purple Yu Mei prefa curcuma as a new flower variety, Anhui agricultural science 42(15) 4607 and 4609.
Zhouxin, Li Zhang Wan, Wangduoping, Liangguang, Li Xia, Chailia 2004. research on effective components of plants of Curcuma of Zingiberaceae, academic report of analysis and test, 23:53-56.
The invention content is as follows:
the invention aims to provide a hybridization method of curcuma plants, which can shorten the fruit development period, improve the fruit setting rate, prolong the pollination time and improve the seed quantity.
The method for crossing curcuma plants of the present invention comprises the following steps:
a. water culture and blooming:
selecting male and female parents from curcuma, selecting male and female parents which can fruit and meet breeding target characters, placing the rhizomes of the male and female parents in a culture chamber, carrying out water culture until inflorescences grow on the rhizomes, wherein the culture solution of the water culture is an improved Hoagland nutrient solution A, and is prepared by adding 50-100mg/L of gamma-aminobutyric acid and 20-30mg/L of glutathione in the Hoagland nutrient solution;
(2) water culture after blooming:
when the bracts at the base of the inflorescence are opened, culturing by using improved Hoagland nutrient solution B;
the improved Hoagland nutrient solution B comprises the following components: adding 20-50mg/L of aminoethoxy vinyl glycine into the improved Hoagland nutrient solution A;
(3) castration
Shearing off anthers of the female parent after the small flowers bloom to castrate;
(4) pollen handling and pollination
Collecting pollen of the male parent, placing the pollen in pollen liquid, stirring uniformly, then pollinating on the head of the female parent, enabling the fruit to ripen and crack, and collecting seeds;
the pollen solution contains beta-nicotinamide mononucleotide 40-50mg/L, polyethylene glycol 6000 5-10g/L, and water in balance.
Preferably, the inflorescence grows from the root stem by water culture and is cultured by using an improved Hoagland nutrient solution B, and the water culture conditions are as follows: 30 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 2000-.
Preferably, the pollen of the male parent is collected by scraping with tweezers or bamboo sticks or dissecting needles, and the pollen of the male parent is collected by pollination, wherein the pollen liquid is dripped on the female parent column head by an injector or sprayed on the female parent florets by a spray can.
Preferably, the pollen concentration in the pollen liquid is 1-2 g/L.
The male parent and the female parent are selected from curcuma, and the specific hybridization combination can be as follows: i: spring and autumn turmeric x queen turmeric, ii: queen curcuma longa x spring and autumn curcuma longa, iii: spring and autumn curcuma longa x curcuma longa or IV: curcuma longa x curcuma longa, v: spring and autumn curcuma longa x Ziyumei Yujin, VI: purple Yumeiji Yujin X Chunqiu Curcuma rhizome, VII: spring and autumn curcuma longa x red cloud curcuma zedoary, VIII: red cloud zedoary, spring and autumn curcuma.
The invention initiates a curcuma species hybridization method in a culture room water culture environment, and the method has the advantages that:
1、
a. one or more plants can be placed in one water culture vessel, the plants can be cultured by pure nutrient solution, or substrates such as sand or ceramsite and the like can be placed at the bottom of the vessel to help to fix the plants, and the nutrient solution is added once a week to keep the liquid level to 1/3-2/3 of the rhizome. The water culture density is 10-30 plants/m2Is more than 10 times of the field cultivation density.
b. The number ratio of the male parent to the female parent is 1: 5 to 1: 10, the pollen collecting amount can meet the requirement of female parent pollination.
c. One culture room can contain one or more female parents, preferably one male parent; if a plurality of male parents are placed in the same culture room, the male parents are separated by gauze to avoid the powder contamination.
Therefore, the management costs of soil preparation, disinfection, plowing, improvement and the like can be saved, the labor intensity is reduced, the land resources are saved, the space utilization rate is improved by water culture, and the problems of inconsistent flowering periods or short overlapping time of different species in the field are solved; realizes closed and standardized management on hybridization environments such as light, temperature and the like, and has higher controllability than field breeding.
2. Before bracts at the base of inflorescence are opened, the water culture nutrient solution is an improved Hoagland nutrient solution A, and the gamma-aminobutyric acid and glutathione in the formula can accelerate the development of fruits and obviously improve the maturing rate. When bracts at the base of inflorescences are opened, the nutrient solution is an improved Hoagland nutrient solution B, and aminoethoxyvinylglycine in the formula can prolong the pollination period of single flowers and the pollination period of inflorescences.
3. The pollen liquid contains beta-nicotinamide mononucleotide, is beneficial to pollen germination and pollen tube elongation, and improves seed collection, and the polyethylene glycol 6000 is an osmotic pressure regulator and prevents pollen from cracking.
Therefore, the invention provides a technical basis for promoting curcuma species crossbreeding work, provides reference significance for developing water culture crossbreeding technologies of other genera in the zingiberaceae, and also provides reference significance for developing water culture crossbreeding technologies of other bulbous plants.
The specific implementation mode is as follows:
the following examples are further illustrative of the present invention and are not intended to be limiting thereof.
Example 1:
(1) hydroponic culture before flowering
The Curcuma longa (Curcuma aromatia) is used as a female parent and is characterized by small and compact plant type, good growth nature and long flowering period, and the Curcuma aromatica (Curcuma aromatia) is used as a male parent and is characterized by strong seed setting capability and is planted in the Panyu, Stamura village farm in Guangzhou city. Placing the rhizomes of the male parent and the female parent in a culture chamber, and carrying out water culture for 20-25 days until inflorescences grow on the rhizomes. The water culture conditions are as follows: the improved Hoagland nutrient solution A has the water culture solution level of 1/3-2/3 of rhizome, 28 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 3000Lux and illumination period of 16 hours/day.
The improved Hoagland nutrient solution A comprises the following components: adding 50mg of gamma-aminobutyric acid and 20mg of glutathione into each liter of Hoagland nutrient solution, and uniformly mixing.
(2) Post-anthesis hydroponic culture
When the bracts at the base of the inflorescence are opened, the nutrient solution is changed into improved Hoagland nutrient solution B, and other culture conditions are not changed.
The improved Hoagland nutrient solution B comprises the following components: aminoethoxyvinylglycine was added to modified Hoagland nutrient solution A to a concentration of 20 mg/L.
(3) Castration
When the small flower is opened, the anther of the female parent is cut off by scissors to castrate.
(4) Pollen handling and pollination
Scraping and collecting pollen of the male parent by using a pair of tweezers, placing the pollen in pollen liquid, uniformly stirring, and then pollinating on the stigmas of the female parent. And marking the hanging plate. The fruits ripen and split in 14-18 days, and the seeds are collected.
The pollen liquid is prepared from beta-nicotinamide mononucleotide, polyethylene glycol 6000 and water by adding beta-nicotinamide mononucleotide 40mg and polyethylene glycol 60005g into 1L of water, and mixing. The pollen concentration in the pollen liquid is 1 g/L.
In this example, 50 pollinations were performed, 16 successful crosses were performed, the maturing rate was 32%, and 157 seeds were harvested.
Example 2:
(1) hydroponic culture before flowering
The method is characterized in that the Curcuma longa (Curcuma aromatica) is used as a female parent, has strong fructification capability, is used as a male parent, has small and compact plant type, good rosette property and long flowering phase, and is planted in the Guangzhou city Panyu Zhongcun farm. Placing the rhizomes of the male parent and the female parent in a culture chamber, carrying out water culture for 20-25 days, and growing inflorescences on the rhizomes. The water culture conditions are as follows: the improved Hoagland nutrient solution A has the water culture solution level of 1/3-2/3 of rhizome, 28 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 3000Lux and illumination period of 16 hours/day.
The improved Hoagland nutrient solution A comprises the following components: 75mg of gamma-aminobutyric acid and 25mg of glutathione are added into each liter of Hoagland nutrient solution and are uniformly mixed.
(2) Post-anthesis hydroponic culture
When the bracts at the base of the inflorescence are opened, the nutrient solution is changed into improved Hoagland nutrient solution B, and other culture conditions are not changed.
The improved Hoagland nutrient solution B comprises the following components: aminoethoxyvinylglycine was added to modified Hoagland nutrient solution A to a concentration of 30 mg/L.
(3) Castration
When the small flower is opened, the anther of the female parent is cut off by scissors to castrate.
(4) Pollen handling and pollination
Scraping and collecting pollen of the male parent by using a pair of tweezers, placing the pollen in pollen liquid, uniformly stirring, and then pollinating on the stigmas of the female parent. And marking the hanging plate. The fruit ripens and cracks in 15-21 days, and the seeds are collected.
The pollen liquid is prepared from beta-nicotinamide mononucleotide, polyethylene glycol 6000 and water, and is prepared by adding 45mg of beta-nicotinamide mononucleotide and 60005g of polyethylene glycol into 1L of water, and mixing uniformly. The pollen concentration in the pollen liquid is 1.5 g/L.
In this example, 50 pollinations were performed, 11 successful crosses were performed, the maturing rate was 22%, and 96 seeds were harvested.
Example 3:
(1) hydroponic culture before flowering
The Curcuma longa (Curcuma aromatia) is used as a female parent and is characterized by small and compact plant type, good rosette and long flowering phase, and the Curcuma longa (Curcuma nankunshanensis) is used as a male parent and is characterized by large plant type, wide and upright leaves and the like which are all planted in the Panyu Zhongcun farm in Guangzhou city. Placing the rhizomes of the male parent and the female parent in a culture chamber, carrying out water culture for 20-25 days, and growing inflorescences on the rhizomes. The water culture conditions are as follows: the improved Hoagland nutrient solution A has the water culture solution level of 1/3-2/3 of rhizome, 28 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 3000Lux and illumination period of 16 hours/day.
The improved Hoagland nutrient solution A comprises the following components: 80mg of gamma-aminobutyric acid and 30mg of glutathione are added into each liter of Hoagland nutrient solution and are uniformly mixed.
(2) After the bracts at the base of inflorescence are opened, the nutrient solution is changed into improved Hoagland nutrient solution B, and other culture conditions are not changed.
The improved Hoagland nutrient solution B comprises the following components: aminoethoxyvinylglycine was added to modified Hoagland nutrient solution A to a concentration of 40 mg/L.
(3) Castration
When the small flower is opened, the anther of the female parent is cut off by scissors to castrate.
(4) Pollen handling and pollination
Scraping and collecting pollen of the male parent by using a pair of tweezers, placing the pollen in pollen liquid, uniformly stirring, and then pollinating on the stigmas of the female parent. And marking the hanging plate. The fruits ripen and crack in 15-22 days, and the seeds are collected.
The pollen liquid is prepared from beta-nicotinamide mononucleotide, polyethylene glycol 6000 and water by adding 45mg of beta-nicotinamide mononucleotide and 60007g of polyethylene glycol into 1L of water and mixing uniformly. The pollen concentration in the pollen liquid is 1.5 g/L.
In this example, 50 plants were pollinated, 10 plants were successfully hybridized, the maturing rate was 20%, and 76 seeds were harvested.
Example 4:
(1) hydroponic culture before flowering
The Curcuma longa (Curcuma nankunshanensis) is used as a female parent and is characterized by large plant type, wide and upright leaves, and Curcuma longa (Curcuma attenuata) is used as a male parent, and the Curcuma longa is characterized by small and compact plant type, good rosette and long flowering phase and is planted in the Panyu Zhongcun farm in Guangzhou city. Placing the rhizomes of the male parent and the female parent in a culture chamber, carrying out water culture for 20-25 days, and growing inflorescences on the rhizomes. The water culture conditions are as follows: the improved Hoagland nutrient solution A has the water culture solution level of 1/3-2/3 of rhizome, 28 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 2000Lux and illumination period of 16 hours/day.
The improved Hoagland nutrient solution comprises the following components: 100mg of gamma-aminobutyric acid and 30mg of glutathione are added into each liter of Hoagland nutrient solution and are uniformly mixed.
(2) Post-anthesis hydroponic culture
When the bracts at the base of the inflorescence are opened, the nutrient solution is changed into improved Hoagland nutrient solution B, and other culture conditions are not changed.
The improved Hoagland nutrient solution B comprises the following components: aminoethoxyvinylglycine was added to modified Hoagland nutrient solution A to a concentration of 50 mg/L.
(3) Castration
When the small flower is opened, the anther of the female parent is cut off by scissors to castrate.
(4) Pollen handling and pollination
Scraping and collecting pollen of the male parent by using a pair of tweezers, placing the pollen in pollen liquid, uniformly stirring, and then pollinating on the stigmas of the female parent. And marking the hanging plate. The fruit ripens and cracks in 20-26 days, and the seeds are collected.
The pollen liquid is prepared from beta-nicotinamide mononucleotide, polyethylene glycol 6000 and water by adding beta-nicotinamide mononucleotide 50mg and polyethylene glycol 600010g into 1L water, and mixing. The pollen concentration in the pollen liquid is 2 g/L.
In this example, 50 plants were pollinated, 6 plants were successfully hybridized, the maturing rate was 12%, and 22 seeds were harvested.
Example 5:
(1) hydroponic culture before flowering
The Curcuma longa (Curcuma aromatia) is used as a female parent and is characterized by small and compact plant type, good growth nature and long flowering period, and the Curcuma aromatica (Curcuma aromatica 'Ziyu') is used as a male parent and is characterized by long flowering branches and strong heat resistance and is planted in the Panyu Zhongcun farm in Guangzhou city. Placing the rhizomes of the male parent and the female parent in a culture chamber, and carrying out water culture for 20-25 days until inflorescences grow on the rhizomes. The water culture conditions are as follows: the improved Hoagland nutrient solution A has the water culture solution level of 1/3-2/3 of rhizome, 28 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 3000Lux and illumination period of 16 hours/day.
The improved Hoagland nutrient solution A comprises the following components: adding 50mg of gamma-aminobutyric acid and 20mg of glutathione into each liter of Hoagland nutrient solution, and uniformly mixing.
(2) Post-anthesis hydroponic culture
When the bracts at the base of the inflorescence are opened, the nutrient solution is changed into improved Hoagland nutrient solution B, and other culture conditions are not changed.
The improved Hoagland nutrient solution B comprises the following components: aminoethoxyvinylglycine was added to modified Hoagland nutrient solution A to a concentration of 20 mg/L.
(3) Castration
When the small flower is opened, the anther of the female parent is cut off by scissors to castrate.
(4) Pollen handling and pollination
Scraping and collecting pollen of the male parent by using a pair of tweezers, placing the pollen in pollen liquid, uniformly stirring, and then pollinating on the stigmas of the female parent. And marking the hanging plate. The fruit ripens and cracks in 15-20 days, and the seeds are collected.
The pollen liquid is prepared from beta-nicotinamide mononucleotide, polyethylene glycol 6000 and water by adding beta-nicotinamide mononucleotide 40mg and polyethylene glycol 60005g into 1L of water, and mixing. The pollen concentration in the pollen liquid is 1 g/L.
In this example, 50 plants were pollinated, 15 plants were successfully crossed, the maturing rate was 30%, and 131 seeds were harvested.
Example 6:
(1) hydroponic culture before flowering
The Curcuma aromatica (Curcuma aromatica 'Ziyu') is used as a female parent and is characterized by long flowering branches and strong heat resistance, and the Curcuma aromatica (Curcuma aromatia) is used as a male parent and is characterized by small and compact plant type, good rosette and long flowering period, and is planted in the Panyu Zhongcun farm in Guangzhou city. Placing the rhizomes of the male parent and the female parent in a culture chamber, carrying out water culture for 20-25 days, and growing inflorescences on the rhizomes. The water culture conditions are as follows: the improved Hoagland nutrient solution A has the water culture solution level of 1/3-2/3 of rhizome, 28 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 3000Lux and illumination period of 16 hours/day.
The improved Hoagland nutrient solution A comprises the following components: 75mg of gamma-aminobutyric acid and 25mg of glutathione are added into each liter of Hoagland nutrient solution and are uniformly mixed.
(2) Post-anthesis hydroponic culture
When the bracts at the base of the inflorescence are opened, the nutrient solution is changed into improved Hoagland nutrient solution B, and other culture conditions are not changed.
The improved Hoagland nutrient solution B comprises the following components: aminoethoxyvinylglycine was added to modified Hoagland nutrient solution A to a concentration of 30 mg/L.
(3) Castration
When the small flower is opened, the anther of the female parent is cut off by scissors to castrate.
(4) Pollen handling and pollination
Scraping and collecting pollen of the male parent by using a pair of tweezers, placing the pollen in pollen liquid, uniformly stirring, and then pollinating on the stigmas of the female parent. And marking the hanging plate. The fruit ripens and cracks in 20-25 days, and the seeds are collected.
The pollen liquid is prepared from beta-nicotinamide mononucleotide, polyethylene glycol 6000 and water, and is prepared by adding 45mg of beta-nicotinamide mononucleotide and 60005g of polyethylene glycol into 1L of water, and mixing uniformly. The pollen concentration in the pollen liquid is 1.5 g/L.
In this example, 50 pollinations were performed, 10 successful crosses were performed, the maturing rate was 20%, and 77 seeds were harvested.
Example 7:
(1) hydroponic culture before flowering
The Curcuma longa (Curcuma aromatia) is used as a female parent and is characterized by small and compact plant type, good rosette property and long flowering phase, and the Curcuma zedoary (Curcuma nankunshanis 'Hongyun') is used as a male parent and is characterized in that leaf masks are purple ribbons which are all planted in the Panyu Zhongcun farm in Guangzhou city. Placing the rhizomes of the male parent and the female parent in a culture chamber, carrying out water culture for 20-25 days, and growing inflorescences on the rhizomes. The water culture conditions are as follows: the improved Hoagland nutrient solution A has the water culture solution level of 1/3-2/3 of rhizome, 28 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 3000Lux and illumination period of 16 hours/day.
The improved Hoagland nutrient solution A comprises the following components: 80mg of gamma-aminobutyric acid and 30mg of glutathione are added into each liter of Hoagland nutrient solution and are uniformly mixed.
(2) After the bracts at the base of inflorescence are opened, the nutrient solution is changed into improved Hoagland nutrient solution B, and other culture conditions are not changed.
The improved Hoagland nutrient solution B comprises the following components: aminoethoxyvinylglycine was added to modified Hoagland nutrient solution A to a concentration of 40 mg/L.
(3) Castration
When the small flower is opened, the anther of the female parent is cut off by scissors to castrate.
(4) Pollen handling and pollination
Scraping and collecting pollen of the male parent by using a pair of tweezers, placing the pollen in pollen liquid, uniformly stirring, and then pollinating on the stigmas of the female parent. And marking the hanging plate. The fruit ripens and cracks in 15-20 days, and the seeds are collected.
The pollen liquid is prepared from beta-nicotinamide mononucleotide, polyethylene glycol 6000 and water by adding 45mg of beta-nicotinamide mononucleotide and 60007g of polyethylene glycol into 1L of water and mixing uniformly. The pollen concentration in the pollen liquid is 1.5 g/L.
In this example, 50 pollinations were performed, 13 successful crosses were performed, the maturing rate was 26%, and 102 seeds were harvested.
Example 8:
(1) hydroponic culture before flowering
The Curcuma zedoary (Curcuma nankunshanensis 'Hongyun') is used as a female parent, is characterized in that a leaf mask is a purple ribbon, and Curcuma longa (Curcuma attenuata) is used as a male parent, is characterized in that the plant type is small and compact, the rosette is good in rosette and long in flowering phase, and is planted in the Panyu Zhongcun farm in Guangzhou city. Placing the rhizomes of the male parent and the female parent in a culture chamber, carrying out water culture for 20-25 days, and growing inflorescences on the rhizomes. The water culture conditions are as follows: the improved Hoagland nutrient solution A has the water culture solution level of 1/3-2/3 of rhizome, 28 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 2000Lux and illumination period of 16 hours/day.
The improved Hoagland nutrient solution comprises the following components: 100mg of gamma-aminobutyric acid and 30mg of glutathione are added into each liter of Hoagland nutrient solution and are uniformly mixed.
(2) Post-anthesis hydroponic culture
When the bracts at the base of the inflorescence are opened, the nutrient solution is changed into improved Hoagland nutrient solution B, and other culture conditions are not changed.
The improved Hoagland nutrient solution B comprises the following components: aminoethoxyvinylglycine was added to modified Hoagland nutrient solution A to a concentration of 50 mg/L.
(3) Castration
When the small flower is opened, the anther of the female parent is cut off by scissors to castrate.
(4) Pollen handling and pollination
Scraping and collecting pollen of the male parent by using a pair of tweezers, placing the pollen in pollen liquid, uniformly stirring, and then pollinating on the stigmas of the female parent. And marking the hanging plate. The fruit ripens and cracks in 20-25 days, and the seeds are collected.
The pollen liquid is prepared from beta-nicotinamide mononucleotide, polyethylene glycol 6000 and water by adding beta-nicotinamide mononucleotide 50mg and polyethylene glycol 600010g into 1L water, and mixing. The pollen concentration in the pollen liquid is 2 g/L.
In this example, 50 plants were pollinated, 7 plants were successfully crossed, the maturing rate was 14%, and 38 plants were harvested.
Comparative test
A,
Before the bracts of inflorescences are opened, the inventor uses improved Hoagland nutrient solution A for water culture, and glutathione is added in the formula, so that fruit setting can be promoted. Compared with the hybridization effect of the Hoagland nutrient solution, the Hoagland nutrient solution A added with gamma-aminobutyric acid and glutathione can shorten the fruit development time and obviously improve the fruit setting rate.
The following specific tests and experimental data provide a more complete understanding of the present invention to those skilled in the art:
gamma-aminobutyric acid is a non-protein amino acid, and physiological effects include nitrogen storage, hormone generation, signal regulation and the like; glutathione is a low molecular weight polypeptide and its physiological effects include storage and transport of reduced sulfur, protein synthesis, and the like. At present, the influence of gamma-aminobutyric acid and glutathione on fructification is not reported, and the influence of the improved Hoagland nutrient solution on fructification is detected by adding gamma-aminobutyric acid and glutathione into the Hoagland nutrient solution.
The method comprises the following specific steps:
(1) placing the rhizomes of the male parent and the female parent in a culture chamber, and carrying out water culture for 20-25 days until inflorescences grow on the rhizomes. The water culture conditions are as follows: the improved Hoagland nutrient solution A (the conventional Hoagland nutrient solution and clear water are used as a contrast), the height of the water culture solution is 1/3-2/3 of the rhizome, 28 +/-1 ℃, the relative humidity is 70-80%, the illumination intensity is 3000Lux, and the illumination period is 16 hours/day.
The improved Hoagland nutrient solution A comprises the following components: adding 50mg of gamma-aminobutyric acid and 20mg of glutathione into each liter of Hoagland nutrient solution, and uniformly mixing.
(2) Post-anthesis hydroponic culture
When the bracts at the base of the inflorescence are opened, the nutrient solution is changed into improved Hoagland nutrient solution B, and other culture conditions are not changed.
The improved Hoagland nutrient solution B comprises the following components: aminoethoxyvinylglycine was added to modified Hoagland nutrient solution A to a concentration of 20 mg/L.
(3) Castration
When the small flower is opened, the anther of the female parent is cut off by scissors to castrate.
(4) Pollen handling and pollination
Scraping and collecting pollen of the male parent by using a pair of tweezers, placing the pollen in pollen liquid, uniformly stirring, and then pollinating on the stigmas of the female parent. And marking the hanging plate. The fruit ripens and cracks, and the seeds are collected.
The pollen liquid is prepared from beta-nicotinamide mononucleotide, polyethylene glycol 6000 and water by adding beta-nicotinamide mononucleotide 40mg and polyethylene glycol 60005g into 1L of water, and mixing. The pollen concentration in the pollen liquid is 1 g/L.
As shown in Table 1, different nutrient solutions have a remarkable influence on the development period of the hydroponic hybrid fruits, the time required for the fruit development is shortest under the culture environment of the improved Hoagland nutrient solution A, the fruits can be harvested and harvested within 15-24 days, and the time required for the fruit development is remarkably increased by using the unmodified Hoagland nutrient solution.
TABLE 1 Effect of different nutritional solutions on the development time of Curcuma fruits
Note: each cross combination was co-pollinated for 20, replicates 3 times. The hybridization combinations (maternal predecessor, same below) were: i: spring and autumn turmeric x Queen turmeric; II: queen curcuma longa x spring and autumn curcuma longa; III: curcuma longa in spring and autumn and Curcuma zedoaria in south Kunshan mountain; IV: curcuma longa L.X.CHUNQIU; v: spring and autumn curcuma longa x Ziyumeiji turmeric; VI: ziyumei Yujin (Curcuma aromatica Salisb) and Chunqiu Curcuma longa (Curcuma longa L.); VII: spring and autumn curcuma longa x red cloud curcuma zedoary; VIII: red cloud zedoary, spring and autumn curcuma. -means no fruit. Different lower case letters after the same column of data represent LSDs0.05The level difference was significant. The following table is the same.
The influence of the nutrient solution on the setting percentage is shown in table 2, when the improved Hoagland nutrient solution A is used, the setting percentage of each hybridization combination is obviously improved, wherein the hybridization performance of the Curcuma longa in spring and autumn and the Curcuma giraldii is the best, and the setting percentage reaches 31.7 percent; the seed setting rate of the improved Hoagland nutrient solution is reduced, which shows that the addition of gamma-aminobutyric acid and glutathione in the Hoagland nutrient solution is beneficial to water culture hybridization seed setting.
TABLE 2 Effect of different nutrient solutions on the hybrid seed set percentage of Curcuma
Note: setting rate (number of set florets/total number of pollinated florets) × 100%. The hybridization combination is as follows: i: spring and autumn turmeric x Queen turmeric; II: queen curcuma longa x spring and autumn curcuma longa; III: curcuma longa in spring and autumn and Curcuma zedoaria in south Kunshan mountain; IV: curcuma longa L.X.CHUNQIU; v: spring and autumn curcuma longa x Ziyumeiji turmeric; VI: ziyumei Yujin (Curcuma aromatica Salisb) and Chunqiu Curcuma longa (Curcuma longa L.); VII: spring and autumn curcuma longa x red cloud curcuma zedoary; VIII: red cloud zedoary, spring and autumn curcuma.
II,
The life of the single-branch inflorescence of the curcuma can reach more than 20 days, bracts are arranged on the inflorescence in a covering tile shape, each bract is provided with a plurality of small flowers, and the blooming time of each small flower is less than 1 day, so that pollination needs to be completed on the day when the small flowers bloom. After bracts at the base of inflorescences are opened, the nutrient solution is changed into an improved Hoagland nutrient solution B, aminoethoxyvinylglycine in the formula is an ethylene synthesis inhibitor, and no report is found about influence of aminoethoxyvinylglycine on pollination at present.
The following specific tests and experimental data provide a more complete understanding of the present invention to those skilled in the art:
(1) placing the rhizomes of the male parent and the female parent in a culture chamber, and carrying out water culture for 20-25 days until inflorescences grow on the rhizomes. The water culture conditions are as follows: the improved Hoagland nutrient solution A has the water culture solution level of 1/3-2/3 of rhizome, 28 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 3000Lux and illumination period of 16 hours/day.
The improved Hoagland nutrient solution A comprises the following components: adding 50mg of gamma-aminobutyric acid and 20mg of glutathione into each liter of Hoagland nutrient solution, and uniformly mixing.
(2) Post-anthesis hydroponic culture
When the bracts at the base of the inflorescence are opened, the nutrient solution is changed into modified Hoagland nutrient solution B (modified Hoagland nutrient solution A is used as a control), and other culture conditions are not changed.
The improved Hoagland nutrient solution B comprises the following components: aminoethoxyvinylglycine was added to modified Hoagland nutrient solution A to a concentration of 20 mg/L.
(3) Castration
When the small flower is opened, the anther of the female parent is cut off by scissors to castrate.
(4) Pollen handling and pollination
Scraping and collecting pollen of the male parent by using a pair of tweezers, placing the pollen in pollen liquid, uniformly stirring, and then pollinating on the stigmas of the female parent. And marking the hanging plate. The fruit ripens and cracks, and the seeds are collected.
Detection of single-flower pollination period: pollinating for 1 time per hour after the small flowers bloom, pollinating for 20 inflorescences each time, pollinating for 2 flowers each inflorescence, pollinating for 40 flowers in total, and repeating for 3 times by taking the maturing rate of more than 5% as the effective pollination period. And (3) detecting the pollination period of the inflorescence: pollinating for 1 time every 10 am, pollinating for 20 inflorescences every time, pollinating for 2 flowers for each inflorescence, pollinating for 40 flowers in total, and repeating for 3 times by taking the maturing rate of more than 5% as the effective pollination period.
The pollen liquid is prepared from beta-nicotinamide mononucleotide, polyethylene glycol 6000 and water by adding beta-nicotinamide mononucleotide 40mg and polyethylene glycol 60005g into 1L of water, and mixing. The pollen concentration in the pollen liquid is 1 g/L.
As shown in table 3, in the reciprocal cross-pollination test with spring and autumn curcuma longa and queen curcuma longa as parents, the single-flower pollination periods of the spring and autumn curcuma longa and queen curcuma longa are 7 hours and 9 hours respectively, the pollination periods of inflorescences are 22 days and 29 days respectively, the single-flower pollination periods of the spring and autumn curcuma longa and queen curcuma longa can be prolonged to 10 hours and 12 hours and the inflorescence pollination period can be prolonged to 31 days and 40 days by using the modified Hoagland nutrient solution B.
TABLE 3 Effect of different nutrient solutions on the pollination phase of Curcuma
Note: detection of single-flower pollination period: pollinating for 1 time per hour after the small flowers bloom, pollinating for 20 inflorescences each time, pollinating for 2 flowers each inflorescence, pollinating for 40 flowers in total, repeating for 3 times with the maturing rate of more than 5% as the effective pollination period, and the p is less than 0.05. And (3) detecting the pollination period of the inflorescence: pollinating for 1 time every 10 am, pollinating for 20 inflorescences every time, pollinating for 2 flowers for each inflorescence, pollinating for 40 flowers in total, using the maturing rate of more than 5% as the effective pollination period, repeating for 3 times, and p is less than 0.05.
III,
The beta-nicotinamide mononucleotide in the pollen liquid is a derivative of nicotinic acid, is a nucleotide with biological activity, is mainly focused on the medical field, is widely involved in the metabolism of human bodies, and is related to immunity and anti-aging. At present, the influence of the beta-nicotinamide mononucleotide on the regulation of plant growth and development is not reported, and the method for treating the pollen by using the beta-nicotinamide mononucleotide can obviously promote the pollen germination and the pollen tube growth and increase the seed number of a single fruit.
The following specific tests and experimental data provide a more complete understanding of the present invention to those skilled in the art:
(1) placing the rhizomes of the male parent and the female parent in a culture chamber, and carrying out water culture for 20-25 days until inflorescences grow on the rhizomes. The water culture conditions are as follows: the improved Hoagland nutrient solution A has the water culture solution level of 1/3-2/3 of rhizome, 28 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 3000Lux and illumination period of 16 hours/day.
The improved Hoagland nutrient solution A comprises the following components: adding 50mg of gamma-aminobutyric acid and 20mg of glutathione into each liter of Hoagland nutrient solution, and uniformly mixing.
(2) Post-anthesis hydroponic culture
When the bracts at the base of the inflorescence are opened, the nutrient solution is changed into modified Hoagland nutrient solution B (taking the modified Hoagland nutrient solution A as a control), and other culture conditions are not changed.
The improved Hoagland nutrient solution B comprises the following components: aminoethoxyvinylglycine was added to modified Hoagland nutrient solution A to a concentration of 20 mg/L.
(3) Castration
When the small flower is opened, the anther of the female parent is cut off by scissors to castrate.
(4) Pollen handling and pollination
Scraping pollen of the male parent with forceps, placing in pollen liquid (if the control is not placed in the pollen liquid, direct pollination), stirring, and pollinating on the female parent stigma. And marking the hanging plate. The fruit ripens and cracks, and the seeds are collected.
The germination test was carried out on pollen treated with the pollen liquid and pollen not treated with the pollen liquid, specifically as follows:
placing two kinds of pollen in germination solution at 28 + -1 deg.C for 3 hr, and inspecting germination condition, wherein the germination solution is: 50g/L of sucrose + 0.1g/L of boric acid + CaCl20.1g/L and the solvent is water. 30 pollen grains were counted per treatment and repeated 3 times.
The pollen liquid is prepared from beta-nicotinamide mononucleotide, polyethylene glycol 6000 and water by adding beta-nicotinamide mononucleotide 40mg and polyethylene glycol 60005g into 1L of water, and mixing. The pollen concentration in the pollen liquid is 1 g/L.
The vitality of the pollen is crucial to the success and failure of cross breeding and the pollen liquid prepared by the invention can promote the pollen germination and the pollen tube growth. The detection results of pollen viability are shown in table 4, the pollen germination rates of the spring and autumn curcuma longa, the queen curcuma longa, the south kunzan curcuma zedoary, the purple-jade-mei-jin and the hongyun curcuma zedoary treated by the pollen liquid are respectively 64.4%, 58.9%, 50.0%, 55.6% and 54.5%, the lengths of the pollen tubes are 511.3 micrometers, 403.2 micrometers, 357.7 micrometers, 386.2 micrometers and 372.9 micrometers, and the pollen germination rates and the pollen tube lengths are all obviously higher than those of untreated pollen.
TABLE 4 Effect of pollen liquid on pollen Germination and pollen tube growth
Note: the germination conditions are that sucrose is 50g/L, boric acid is 0.1g/L and CaCl20.1g/L, 28 +/-1 ℃ and 3 h. 30 pollen grains were counted per treatment, repeated 3 times, p<0.05. The pollen control group is pollen not treated with pollen solution; the pollen treatment group is pollen treated with pollen liquid.
Further, the influence of the pollen liquid on the seeds was examined by using a hybridization test, and it can be seen from the results in table 5 that the pollination of the stigma with pollen liquid (treatment group) significantly increased the number of seeds of a single fruit, wherein the hybridization of spring and autumn curcuma longa x curcuma longa was the best and the number of seeds of a single fruit was 10.
TABLE 5 influence of pollen treatment on the number of hybrid seeds
Note: 10 fruits were counted per treatment, repeated 3 times with p < 0.05. The pollen control group is pollen not treated with pollen solution; the pollen treatment group is pollen treated with pollen liquid. The hybridization combination is as follows: i: spring and autumn turmeric x Queen turmeric; II: queen curcuma longa x spring and autumn curcuma longa; III: curcuma longa in spring and autumn and Curcuma zedoaria in south Kunshan mountain; IV: curcuma longa L.X.CHUNQIU; v: spring and autumn curcuma longa x Ziyumeiji turmeric; VI: ziyumei Yujin (Curcuma aromatica Salisb) and Chunqiu Curcuma longa (Curcuma longa L.); VII: spring and autumn curcuma longa x red cloud curcuma zedoary; VIII: red cloud zedoary, spring and autumn curcuma.
Claims (5)
1. A method of crossing plants of the genus curcuma, comprising the steps of:
a. water culture and blooming:
selecting male and female parents from curcuma, selecting male and female parents which can fruit and meet breeding target characters, placing the rhizomes of the male and female parents in a culture chamber, carrying out water culture until inflorescences grow on the rhizomes, wherein the culture solution of the water culture is an improved Hoagland nutrient solution A, and is prepared by adding 50-100mg/L of gamma-aminobutyric acid and 20-30mg/L of glutathione in the Hoagland nutrient solution;
(2) water culture after blooming:
when the bracts at the base of the inflorescence are opened, culturing by using improved Hoagland nutrient solution B;
the improved Hoagland nutrient solution B comprises the following components: adding amino ethoxy vinyl glycine into the improved Hoagland nutrient solution A to 20-50 mg/L;
(3) castration
Shearing off anthers of the female parent after the small flowers bloom to castrate;
(4) pollen handling and pollination
Collecting pollen of the male parent, placing the pollen in pollen liquid, stirring uniformly, then pollinating on the head of the female parent, enabling the fruit to ripen and crack, and collecting seeds;
the pollen solution contains beta-nicotinamide mononucleotide 40-50mg/L, polyethylene glycol 6000 5-10g/L, and water in balance.
2. The hybridization method according to claim 1, wherein said water culture to grow inflorescence on the root-like stem and modified Hoagland nutrient solution B is used for culture under the following water culture conditions: 30 +/-1 ℃, relative humidity of 70-80%, illumination intensity of 2000-.
3. The hybridization method according to claim 1, wherein the pollen of the male parent is collected by scraping with tweezers, bamboo sticks or dissecting needles, and the pollination is performed by dripping the pollen liquid on the female parent through a syringe or spraying the pollen liquid on the female parent florets through a watering pot.
4. The hybridization method according to claim 1, wherein the parent and the female parent are selected from the genus curcuma, and the specific hybridization combination is: i: spring and autumn turmeric x queen turmeric, ii: queen curcuma longa x spring and autumn curcuma longa, iii: spring and autumn curcuma longa x curcuma longa, IV: curcuma longa x curcuma longa, v: spring and autumn curcuma longa x Ziyumei Yujin, VI: purple Yumeiji Yujin X Chunqiu Curcuma rhizome, VII: spring and autumn curcuma longa x red cloud curcuma zedoary, VIII: red cloud zedoary, spring and autumn curcuma.
5. The hybridization method according to claim 1, wherein the concentration of pollen in the pollen liquid is 1 to 2 g/L.
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| CN101536641A (en) * | 2009-04-15 | 2009-09-23 | 仲恺农业工程学院 | Guangxi zedoary florescence control method |
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| CN101536641A (en) * | 2009-04-15 | 2009-09-23 | 仲恺农业工程学院 | Guangxi zedoary florescence control method |
| CN101536663A (en) * | 2009-04-15 | 2009-09-23 | 仲恺农业工程学院 | Guangxi zedoary aquiculturing and blooming method |
| CN104285537A (en) * | 2014-09-19 | 2015-01-21 | 广州普邦园林股份有限公司 | Method for increasing flowering rate of rootstalks of Hongyun curcuma zedoary cultured in water |
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