阅读说明：本技术 使用切叶蜂属的蜂的莴苣属植物种子的生产方法 (Method for producing seed of Lactuca plant using wasp of genus Cerebrophora ) 是由 堀内慎吾 铃木隆夫 泉田敦 于 2018-12-05 设计创作，主要内容包括：本发明涉及莴苣属植物的种子的生产方法,所述方法包括：作为花粉传播手段,使用切叶蜂属的蜂,并且,除了花粉亲本莴苣属植物和种子亲本莴苣属植物之外,还使用下述植物种作为辅助蜜源植物,进行莴苣属植物的交配,所述植物种是切叶蜂属的蜂具有访花偏好性、并且该访花偏好性等于或低于莴苣属植物的植物种。根据本发明,能够使用切叶蜂属的蜂作为花粉传播手段,并且与以往相比高效且经济性地生产莴苣属植物的种子。(The present invention relates to a method for producing seeds of a plant of the genus lactuca, said method comprising: as a pollen transmission means, bees of the genus Apis are used, and mating of plants of the genus Lactuca is carried out using, as a co-nectar source plant, in addition to the pollen parent Lactuca plant and the seed parent Lactuca plant, a plant species in which bees of the genus Apis have a floral preference and which floral preference is equal to or lower than that of the Lactuca plant. According to the present invention, it is possible to produce seeds of a plant of the genus lactuca more efficiently and economically than before using bees of the genus aphelesia as a pollen propagation means.)

1. A method of producing seed of a plant of the genus lactuca, the method comprising:

as a pollen transmission means, a bee of genus Cucubaria is used, and,

mating of the lactuca plants is carried out using, in addition to the pollen parent lactuca plant and the seed parent lactuca plant, as a honey-assist source plant, a plant species of the genus lactuca which has a floral preference for bees of the genus aphania and which is equal to or lower than that of the lactuca plant.

2. The method of claim 1, wherein the lettuce seeds are hybrid first generation lettuce seeds (F1 seeds).

3. The method as claimed in claim 1 or 2, wherein the plant of the genus Lactuca is Lactuca sativa (Lactuca sativa).

4. The method of any one of claims 1 to 3, wherein the bees of the genus Cerebrophora are Medicago cerifera.

5. The method of any one of claims 1 to 4, wherein the helper honey plants have a flowering habit such that they also flower when the lettuce plant closes.

6. The method according to any one of claims 1 to 5, wherein 1 or more genera selected from the group consisting of plants of the genus Sphaeranthus, Coriandrum, Hyssopus, Mentha, Nepeta, Potentilla, Salvia, erigeron, Foeniculum, Helichrysum, Portulaca, and Phalaenopsis are used as the auxiliary nectarine plant.

7. The method according to any one of claims 1 to 6, wherein 1 or more selected from the group consisting of chamomile (Anthemitinaria), coriander (Coriandrum sativum), hyssop officinalis (Hyssopus officinalis), Mentha officinalis (Monarda didyma), Schizonepeta tenuifolia (Nepeta cataria), Potentilla pulchella (Pentappets Phoenica), Salvia officinalis (Salvia azurea), Callerbiviea fraxinifolia (Erigeeron karvinskianus), Foeniculum vulgare (Foeniculumvulgare), Matricaria chamomilla (Melampium paludousum), Portulaca grandiflora (Portulaca grandiflora), and corydalis edulis (Torremia officinalis) are used as the auxiliary nectaricus plants.

8. The method according to any one of claims 1 to 7, wherein 1 or more selected from the group consisting of Calibeiya, fennel, Matricaria, Portulaca grandiflora, and corydalis impatiens are used as auxiliary nectarine-derived plants.

9. The method according to any one of claims 1 to 8, wherein 1 or 2 selected from the group consisting of Matricaria and corydalis impatiens are used as auxiliary honey-source plants.

10. The method according to any one of claims 1 to 9, wherein, as the auxiliary nectar source plant, a melastoma grandis is used.

11. The method of any one of claims 1 to 10, wherein the pollen parent Lactuca plant is a male fertile line and the seed parent Lactuca plant is a male sterile line.

12. A method for mating Lactuca plants, wherein bees belonging to genus Cerasifera are used as pollen transmission means,

the method is characterized in that, as an auxiliary honey-source plant, bees of the genus Ceriporia have a floral preference and the floral preference is equal to or lower than that of a plant of the genus Lactuca are used.

13. The method according to claim 12, wherein the mating of the Lactuca plants is aimed at obtaining seeds of the first hybrid generation (F1) of the Lactuca plants.

14. The method as claimed in claim 12 or 13, wherein the plant of the genus Lactuca is Lactuca sativa (Lactuca sativa).

15. The method of any one of claims 12 to 14, wherein the bees of the genus Cerebrophora are lucerne Cerebrophora.

16. The method of any one of claims 12 to 15, wherein the auxiliary honey-derived plant has a flowering habit such that it also flowers when the lettuce plant closes.

Technical Field

The present invention relates to a method for producing seeds of a plant of the genus Lactuca with high efficiency using a bee of the genus Cerebrophora as a pollen transmission means. More specifically, the present invention relates to a method for producing seeds of a lettuce plant that uses a bee of the genus Cerebrophora as a pollen transmission means and uses a honey-assisting plant in addition to a pollen parent lettuce plant and a seed parent lettuce plant. More specifically, the present invention relates to a method for producing a first-generation hybrid seed (F1 seed) of lettuce.

Background

Among plant varieties, there are fixed-seed and hybrid first-generation (hereinafter sometimes simply referred to as "F1") varieties, and among major crops, F1 varieties are prevalent.

The F1 variety has the great advantages of vigorous growth, fast growth, improved yield capability and the like due to heterosis (heterosis). Further, since the F1 cultivar has vigorous growth, it is expected to improve environmental suitability such as tolerance to disease and pest insects, cold resistance and heat resistance. In addition, since the genotype of the F1 variety is heterozygous and the same genotype, the phenotype shows extremely high uniformity, and the marketability of the product is improved. In addition, since the parents of the F1 variety can integrate useful traits governed by dominant genes, rapid breeding can be achieved. The F1 variety dominates the main crop cultivars due to the advantages described above.

Lactuca, especially lettuce (lettuce)Lactuca sativa) Vegetables are produced in various countries all over the world and have a very large market size, and F1 is strongly demanded to be bred. The present inventors have succeeded in developing cytoplasmic male sterile lettuce which is essential for the development of the lettuce variety F1, and have shown that F1 seeds can be produced even in lettuce (international publication No. WO2007/049730a (patent document 1)).

The lettuce seeds F1 were collected by hand mating, but the lettuce flowers only several hours in the morning, and therefore the handling efficiency was poor, and the number of seeds obtained by mating 1 flower was as small as about 10. Therefore, seed collection by manual mating is economically inefficient, and development of an alternative, economically efficient seed collection method is desired. Although it is desired to develop an economical mating and seed-collecting method using pollinating insects as carried out in other crops, it is necessary to use pollinating insects other than western bees because the lettuce is not visited by the western bees (Apis mellifera) generally known as pollinating insects.

Therefore, in order to solve the above-described problems in lettuce, various pollinating insects have been studied so far for their use.

For example, japanese patent No. 3635036 (patent document 2) discloses a method for collecting F1 seeds of lettuce by using lasioglossoum villosulum. However, the number of life-saving venomous bees is small, and even if they are successfully captured, the proliferation method thereof has not been disclosed. Further, according to the data described in this patent document, the seed collection efficiency is also extremely low, and it is considered difficult to use for commercial seed collection from the economical viewpoint, because 330 grains per 1 plant (about 0.33g because the weight of one thousand grains of lettuce is about 1 g). In addition, channelan palustris is not marketed as yet and is often difficult to obtain, and is therefore considered difficult to use in commercial breedings that require large numbers of pollinating insects.

Further, japanese patent No. 4471983 (patent document 3) discloses a method for collecting F1 seeds of lettuce by lucilia sericata (luciliaceae). However, according to the pollination ability test result using lucilia sericata described in this patent document, the number of fruit set is only 245 grains (about 0.245g) in total from 2 plants, and it is considered difficult to use for commercial seed collection from the economical point of view as in the case of patent document 2. In addition, since flies do not home, closed facilities are required for seed collection. In commercial seed collection, it is desirable to be able to produce seeds in an open nursery that is inexpensive to produce. Therefore, the method of patent document 2 may increase the cost.

Japanese patent No. 5627894 (patent document 4) discloses a method of utilizing Bactrocera dorsalis of the order Diptera of the family Liriomyidae (see below)Calliphora vomitaria) Red head blowfly (A. rubra.), (Calliphora erythrocephala) Or Kaiserda (a)Lucilia Caesar) To collect lettuce F1 seeds. According to the method of the patent document, F1 seeds were obtained in an amount of 1.67 to 11.25g per 1 lettuce.

However, as described in paragraph 0040 of this patent document, Diptera (Diptera) is not an insect that is constantly pollinated to lettuce flowers, and is also not known to feed on lettuce nectar. Patent literature only describes that an excessive number of diptera are introduced into a closed environment and then act as pollen pollinators for lettuce. In addition, according to the examination document (written document by the applicant) of the examination process of the patent document, it is explained that diptera is not specifically attracted by the flowers of lettuce, but by using a large amount of diptera, at least a part of which probabilistically falls to the corolla and comes into contact with pollen, thereby achieving pollination. Therefore, this method is inferior in pollination efficiency compared with pollinating insects that actively visit flowers for ingestion, and therefore an excessive number of diptera must be introduced. Further, as in the case of patent document 3, since flies do not have homing properties, seed collection requires a closed facility. Therefore, there is a possibility that the production cost of the seeds increases.

Japanese patent application laid-open No. 2009-539409 (patent document 5) (International publication WO 2007/146420A) discloses the use of wasps of the genus Cucubeba, specifically alfalfa Cucubeba: (Megachile rotundata) To collect F1 seeds of lettuce. Among them, a method for attracting alfalfa cutting bees and allowing the alfalfa cutting bees to visit lettuce by cultivating alfalfa close to lettuce is disclosed. International publication No. WO2009/086173a (patent document 6) discloses a method for collecting F1 seeds of lettuce by using aphelenchoides pteronyssinus, specifically alfalfa aphelenchoides pteronyssinus, as in patent document 5.

However, the examples of patent document 5 disclose only: an average of 5.6 seeds were obtained for 24 flowers with male sterility (patent document 5, paragraph 0065); and, 13 different female alfalfa cutting bees were observed every 2 minutes, and it was found that 1 minute visit was average to 7.1 flowers (patent document 5, paragraph 0096). Patent document 5 does not disclose specific and quantitative seed collection amounts, for example, the yield of F1 lettuce seeds per 1 plant and the yield of F1 lettuce seeds per unit plot area. These points are also the same for patent document 6.

Therefore, it is difficult for those skilled in the art who have read patent documents 5 and 6 to conceive F1 seeds capable of collecting lettuce at a commercial level according to the method disclosed therein.

Disclosure of Invention

Problems to be solved by the invention

The purpose of the present invention is to provide a method for producing seeds of a lettuce plant efficiently and economically compared to the conventional method, using a bee of the genus Cerebrophora as a pollen propagation means.

Means for solving the problems

The alfalfa cutting wasp is the most widely used pollinator in alfalfa breeds in north america (reference 1(w.p. kemp et al, (2000)), p 904). In addition, the alfalfa leaf cutting bees are also known as pollinators for crops other than alfalfa, specifically, blueberry, Borage (Borage), Canola (Canola), carrot, cranberry, crow flower (crow vent), hair vetch (Hairy vent), mint, onion, and corydalis saxicola (Sweet vent) (reference 2(Eric Mader et al, (2010)), p10, table 1.2). Further, the alfalfa leaf cutting wasps have excellent characteristics as pollinators (assimilation, continuous flower pollination, high pollination efficiency, easy management, and nesting property adapted to artificial environment) (reference 3 (male with rosin light, "insect utilization science series 7" (1996)), p 99).

The pollinating efficiency is high in all species because the wasps such as alfalfa cutting wasps need to collect pollen and honey for offspring. On the other hand, in the case of flies, for example, the parent flies only lay eggs, and the larvae of the offspring feed on their own from putrefactive substances and the like, so the parent flies only need to collect honey as their own active energy source and nutrient source for egg production.

In the flower bees, since the parent bees need to collect and provide the offspring as a nutrient source until the imagoes become, 1 offspring needs a large amount of honey and pollen. Especially for solitary wasps, the next generation of breeding depends on the labor of the parent bees and needs to be achieved in a short time. Therefore, in the case of the wasps, the amount of labor of 1 individual is remarkably remarkable, and it is said that the amount of labor is 20 to 80 times that of social bees.

As described above, the ability of alfalfa leaf cutting bees as pollinators is very high, and it is considered promising to replace bees as pollinators in various crops. In addition, alfalfa breedings are primarily conducted in the western united states, with alfalfa cutting bees produced in canada and imported into the united states in large quantities each year. The main breeding ground for lettuce is also the western united states, and it is therefore also a great advantage to be able to easily obtain a large number of alfalfa cutting bees.

Alfalfa secatella is reported to be a europaea species with a strong preference for leguminous plants while visiting a wide range of compositae plants (reference 4(Dave Goulson, annu. rev. ecol. evol. syst.34:1-26(2003)), p 3).

In the method described in the aforementioned patent document 5 (japanese patent application laid-open No. 2009-539409), alfalfa is used as an attractant plant. However, alfalfa is higher than lettuce in terms of its floral interview preference, and therefore, in the case of mixed lettuce and alfalfa, the floral interview of alfalfa cutting bees will be concentrated on alfalfa. As a result, the efficiency of powdering in lettuce is not improved, and seed collection has to be restricted.

Patent document 5 (paragraph 0048) also describes that alfalfa flowers can be cut off after lettuce flowers so that alfalfa leaf cutting bees can be concentrated on lettuce.

Here, however, cutting out alfalfa flowers may cause significant problems as described below.

That is, lettuce flowers have a special flowering habit that flowers bloom in the early morning and close flowers several hours later, so that, in the case of alfalfa flowers being cut, the seed garden loses all honey resources after lettuce flowers close. On the other hand, alfalfa leaf cutting bees will continue to feed during the day when the temperature is above about 25 ℃. Therefore, it repeats active flights aimed at finding honey sources, and most individuals are physically consumptive, and if the honey sources are insufficient, starvation is caused within the day. Thus, as previously described, alfalfa leaf cutting bees will gradually starve to death when no honey source is present in the seed nursery after the lettuce has closed due to the cut flowers of alfalfa.

In addition, the operation of cutting all alfalfa flowers in the seed collection nursery every time the alfalfa leaf cutting bees are stocked is complicated, and requires a very large labor cost.

As described above, lettuce has a special flowering habit that flowers only in the morning for several hours, while many pollinating insects have a day-to-day periodicity of the type that performs feeding activities over a long diurnal period, as described in patent document 2 (paragraph 0006).

Therefore, it is difficult to focus pollinating insects on a particular plant for only a short time of morning feeding.

In addition, patent document 5 describes that, in addition to alfalfa being used as an attractant plant, a plant extract product containing a plant composition for attracting alfalfa leaf cutting bees can be placed in the vicinity of lettuce or placed in contact with lettuce. However, such a report of an attractant extract product effective for alfalfa leaf cutting bees is not known in practice, and patent document 5 also does not specifically disclose it. Therefore, it is difficult for those skilled in the art to easily obtain and use such an attractive extraction product.

Based on the above-described circumstances, the inventors of the present application have made intensive studies repeatedly.

The inventors of the present application have conducted the following studies: in producing seeds of a lettuce plant, a pollen parent lettuce plant and a seed parent lettuce plant are prepared in a seed nursery of the lettuce plant, and mating of the lettuce plant is performed using a bee of the genus Ceratoptera as a pollen propagation means. At this time, in order to prevent bees from becoming hungry when the lettuce plant closes, honey-assisting plants were prepared together, and mating of the lettuce plant was performed using the bees of the genus Cerebrophora as a pollen transmission means. As the auxiliary honey source plant, a species of a bee of the genus Cerebrophora having a floral preference which is equal to or lower than that of the genus Lactuca is used. As a result, by preparing the auxiliary honey-source plant, it is possible to substantially suppress the hornet from becoming hungry, and thereby, it is possible to efficiently mate the lettuce plant and successfully produce seeds more efficiently.

In the present invention, the above findings are used as a basis.

That is, according to the present invention, the following invention can be provided.

A method of < 1 > producing seed of a plant of the genus Lactuca, the method comprising:

as a pollen transmission means, a bee of genus Cucubaria is used, and,

mating of the lactuca plants is carried out using, in addition to the pollen parent lactuca plant and the seed parent lactuca plant, as a honey-assist source plant, a plant species of the genus lactuca which has a floral preference for bees of the genus aphania and which is equal to or lower than that of the lactuca plant.

< 2 > the method as described above < 1 >, wherein the seed of the Lactuca plant is a hybrid first-generation seed of the Lactuca plant (F1 seed).

< 3 > the method as defined above < 1 > or < 2 > wherein the plant of the genus Lactuca is Lactuca sativa (Lactuca sativa L.) (Lactuca sativa)。

< 4 > the method according to any one of the above < 1 > -to < 3 >, wherein the wasp of genus Cucubeba is a Medicago Cucubeba.

< 5 > the method according to any one of the above < 1 > -4 >, wherein the honey-assisted plant has a flowering habit that the lettuce plant flowers also when closing flowers.

< 6 > the method as defined in any of the above < 1 > - < 5 >, wherein as the auxiliary honey-derived plant, 1 or more genera selected from the group consisting of a plant of the genus chamomile (Anthemi), a plant of the genus coriander (Coriandrum), a plant of the genus Hyssopus (Hyssopus), a plant of the genus peppermint (Monarda), a plant of the genus Nepeta, a plant of the genus Pentium (Pentapaeus), a plant of the genus Salvia officinalis (Salvia), a plant of the genus Erigeron (Erigeeron), a plant of the genus Foeniculum (Foenicum), a plant of the genus Melamopsis (Melampidium), a plant of the genus Portulaca (Portulaca), and a plant of the genus Iris japonica (Torenia) are used.

< 7 > the method as defined in any of the above < 1 > - < 6 >, wherein a plant selected from among chamomile (F) is used as an auxiliary honey sourceAnthemis tinctoria) Coriander (f) (coriander)Coriandrum sativum) Hyssop (hyssopus officinalis), american mint (Monarda didyma), schizonepeta (catnip)Nepeta cataria) Noon time flower (1)Pentapetes phoenicea) Blue sage (1)Salvia azurea) "Kahlebi Feihu (a)Erigeron karvinskianus) Fennel fruit (fennel)Foeniculum vulgare) "Meilanju" (a. Merrill.), (Melampodium paludosum) Herba Portulacae (herba Portulacae)Portulaca grandiflora) And Xiayu (Xiayu)Torenia fournieri) 1 or more of the group.

< 8 > the method according to any one of the above < 1 > -7 >, wherein 1 or more selected from the group consisting of Calibeibia, Foeniculum vulgare, Matricaria chamomilla, Portulaca grandiflora, and corydalis impatiens are used as the auxiliary honey source plant.

< 9 > the method as defined in any one of the above < 1 > -to < 8 >, wherein 1 or 2 selected from the group consisting of Matricaria chamomilla and corydalis impatiens are used as auxiliary honey-source plants.

< 10 > the method as defined in any one of the above < 1 > -to < 9 >, wherein, as the auxiliary nectar source plant, a chrysanthemum morifolium is used.

< 11 > the method as defined in any of the above < 1 > - < 10 >, wherein the pollen parent lettuce plant is a male fertile line and the seed parent lettuce plant is a male sterile line.

< 12 > A method for mating Lactuca plants, wherein bees belonging to genus Cerasifera are used as pollen transmission means,

the method is characterized in that, as an auxiliary honey-source plant, bees of the genus Ceriporia have a floral preference and the floral preference is equal to or lower than that of a plant of the genus Lactuca are used.

< 13 > the method of < 12 > as described above, wherein the purpose of mating lettuce plants is to obtain seeds of the first hybrid generation (F1) of lettuce plants.

< 14 > the method < 12 > or < 13 > as described above, wherein the plant of the genus Lactuca is Lactuca sativa (Lactuca sativa L.) (Lactuca sativa)。

< 15 > the method according to any one of the above < 12 > -to < 14 >, wherein the wasp of genus Cucubeba is a Medicago Cucubeba.

< 16 > the method as defined in any of < 12 > - < 15 > above, wherein the honey-assisted plant has a flowering habit that the lettuce flowers also when closing flowers.

Effects of the invention

According to the present invention, seeds of a lactuca plant, which have been difficult to be produced efficiently by pollinating insects until now, can be produced more efficiently than in the past by breeding an auxiliary honey-source plant in a seed nursery of a lactuca plant and using a bee of the genus Cerebrophora as a pollen propagation means. In addition, the method of the present invention can significantly reduce the labor and the operation for producing seeds of a lettuce plant, and can also significantly reduce the production cost.

Drawings

FIG. 1 is a graph showing the results of the investigation of the frequency of flower visits between different plant species of example 3.

FIG. 2 shows the configuration of the seed parent, pollen parent, and auxiliary nectar plant (alfalfa or meadow chrysanthemum) in the greenhouse of example 4 (the "nectar plant" in the figure).

FIG. 3 shows the configuration of the seed parent, pollen parent, and auxiliary honey plant (corydalis impatiens) (in the figure, "honey plant") in the greenhouse in example 5.

Detailed Description

The present invention will be described in detail below.

The method for producing seeds of a lactuca plant according to the present invention is as described above, and comprises: as a pollen transmission means, bees of the genus Apis are used, and mating of plants of the genus Lactuca is carried out using, as a co-nectar source plant, in addition to the pollen parent Lactuca plant and the seed parent Lactuca plant, a plant species in which bees of the genus Apis have a floral preference and which floral preference is equal to or lower than that of the Lactuca plant.

In the present invention, seed production of a plant of the genus Lactuca is performed. Here, the production of seeds means as follows: the seed is produced by mating a lettuce plant which becomes a pollen parent with a lettuce plant which becomes a seed parent, that is, by receiving pollen of the pollen parent in the seed parent and collecting the seed obtained by cultivating the seed parent.

Examples of the plant of the genus Lactuca include Lactuca sativa (L.) (Lactuca sativa) Poisonous lettuce (Du lettuce)Lactuca serriola)、L.aculeate、L.scarioloides、L.azerbaijanica、L.georgica、L.dregeanaAltai lettuce (see below)L.altaica) Caulis et folium Lactucae sativaeL.saligna) Lettuce with spiny hair (L.virosa) Lettuce (see below) as Mongolian medicineL.tatarica) Lettuce (Lactuca sativa L.)L.indica) Or isL.debilisAnd the like. Lettuce (Lactuca sativa L.) may also be included in the genus LactucaLactuca sativa) A hybrid formed by cell fusion with a plant species of the genus Lactuca other than the above, or a hybrid formed by interspecies crossing. Preferably, the lettuce plant is at least one selected from the group of species listed above, and more preferably lettuce (see below)Lactuca sativa)。

In the method of the present invention, a pollen propagation means for transporting pollen from a pollen parent plant to pollinate a seed parent plant is used by a flowering insect. As such a flowering insect, a bee of the genus serenoa, which is known to perform flowering on a plant of the genus lactuca, is used. Such a bee of genus Apis is not particularly limited, and specific examples thereof include alfalfa cutting bees (Megachile rotundata)、M.latimanus(reference 5),M.brevis(reference 6),M.montivaga(reference 7),M.onobrychidis(reference 8),M.dentitarsus(reference 9),M.relativa(reference 10). These may be used alone or in combination of two or more. In the present invention, alfalfa secatella is preferable as the bee of the genus secatella used as a flower-visiting insect, from the viewpoint of being easily available on the market.

The seed of the lettuce plant produced in the present invention is preferably a hybrid first generation seed (F1 seed). Here, the first filial generation (F1) is a sub-generation generated by crossing between parents having each gene in a certain allele in a homozygous form.

In the present invention, the seed parent means a parent line of the seed produced by the method of the present invention, and is a parent plant which is treated (pollen is flushed away with water, etc.) or cultivated (i.e., cultivated to have male sterility) so as not to be pollinated with self-pollen having insemination ability. When the seed produced in the present invention is F1 seed, the seed parent is preferably a line (male sterile line) which is cultivated so that pollen having insemination ability cannot be formed by itself. Specifically, for example, a seed parent line can be obtained by backcrossing a line having male sterile cytoplasm as a female parent and a non-recurrent parent and a target line confirmed to have no fertility restoring factor as a recurrent parent, and by replacing the cytoplasm of the target line with the male sterile cytoplasm, thereby imparting male sterility.

In the present invention, the seed parent lactuca plant means a lactuca plant which becomes a seed parent. The seed parent lactuca plant is preferably a male sterile line, more preferably a Cytoplasmic Male Sterile (CMS) line. Examples of such a CMS strain include the strain described in patent document 1.

In the present invention, the pollen parent refers to a parent strain of the seed produced by the method of the present invention, and is a strain to which pollen is supplied. When the seeds produced in the present invention are F1 seeds, the pollen parent is a male fertile line.

In the present invention, the pollen parent lactuca plant means a lactuca plant which is a pollen parent. The pollen parent lactuca plant is preferably a male fertile line.

Generally, in a seed nursery, in order to make pollination of pollen uniform, it is desirable that pollen parents (preferably male fertile lines) and seed parents (preferably male sterile lines) are planted in rows or alternately on the basis of the ratio of the pollen parents to the seed parents.

In the present invention, helper nectar source plants are used in addition to the pollen parent plant and the seed parent plant.

Lettuce flowers, such as lettuce flowers, have a flowering habit that flowers open in the early morning hours and close flowers after hours. Therefore, in order to prevent bees as a means of pollen propagation from starving due to insufficient honey sources after the lettuce is closed, auxiliary honey-source plants are used in the present invention. Auxiliary honey plants are prepared around the hive of bees or around the lettuce plant as the pollen parent and/or the seed parent, and are allowed to bloom there as honey sources.

Therefore, the auxiliary honey source plant in the present invention is a plant species in which bees of the genus Cerebrophora have a floral preference, and the floral preference is equal to or lower than that of the genus Lactuca. Even when the floral preference is equal to that of lettuce, seed production of lettuce using bees of the genus Cerebrophora can be efficiently performed as long as the lettuce can be a honey source when closing flowers. It can be confirmed whether or not the floral preference of bees is equal to or lower than that of the lettuce plant species by, for example, the method described in example 1 described later. The auxiliary honey source plant is preferably a plant species of the alfalfa secatella having a floral preference which is equal to or lower than lettuce.

The auxiliary honey source plant is more preferably a plant species in which bees of the genus Cerebrophora have a floral preference lower than that of the genus Lactuca, and still more preferably a plant species in which Medicago sativa has a floral preference lower than that of Lactuca.

As the plant species that can be used as the auxiliary nectariferous plant, a plant species having not only the aforementioned flower-preferred property but also a flowering habit that flowers when lettuce closes. Specifically, the plant species that can be used as the auxiliary nectar source plant is preferably a plant species that has excellent flowering-sustaining properties as shown in example 1 described later. Further, a plant species that can be used as an auxiliary nectar source plant is advantageous as a plant species having excellent heat tolerance. The heatstroke resistance referred to herein can be evaluated by the method described in example 1, which will be described later.

In the present invention, the plant capable of serving as the auxiliary nectar source plant is not particularly limited, and examples thereof include plants of the genera chamomilla, coriander, hyssop, monarda, nepeta, noontia, salvia, erigeron, anisum, merantium, portulaca, and phalaenopsis. According to a preferred embodiment of the present invention, 1 or more species selected from the above-mentioned genera can be used as the auxiliary nectariferous plant. In the present invention, among the plants of the above-mentioned genus, those having not only the aforementioned flower-preferred property but also a flowering habit of flowering lettuce when closing the flower are preferably used.

According to a further preferred embodiment of the present invention, as the plant capable of serving as the auxiliary nectar source plant, 1 or more species selected from plants of erigeron, fennel, meadowsweet, purslane and phalaenopsis can be used, and more preferably, 1 or 2 species selected from plants of meadowsweet and phalaenopsis are used, and particularly, a plant of meadowsweet is used.

According to a preferred embodiment of the present invention, the plant species which can be used as the auxiliary nectariferous plant includes chamomile (Chamomilla recutita, Chamomilla recutiAnthemis tinctoria) Coriander (f) (coriander)Coriandrum sativum) Achyranthes bidentata (Hyssopus officinalis) American mint (B)Monarda didyma) Schizonepeta (A) and (B)Nepeta cataria) Noon time flower (1)Pentapetes phoenicea) Blue sage (1)Salvia azurea) "Kahlebi Feihu (a)Erigeron karvinskianus) Fennel fruit (fennel)Foeniculum vulgare) "Meilanju" (a. Merrill.), (Melampodium paludosum) Herba Portulacae (herba Portulacae)Portulaca grandiflora) And Xiayu (Xiayu)Torenia fournieri). According to a preferred embodiment of the present invention, 1 or more species selected from the above-listed plant species can be used as the auxiliary nectariferous plant.

According to a more preferred embodiment of the present invention, 1 or more species selected from the group consisting of California, fennel, Matricaria, Portulaca grandiflora, and corydalis impatiens are used as the auxiliary honey source plant in the method of the present invention. More preferably, 1 or 2 species selected from the group consisting of cymbidium and corydalis impatiens are used as the auxiliary honey source plant, and cymbidium is particularly preferably used.

It is desirable for the auxiliary honey-derived plants to appropriately adjust the planting period so as to match the flowering period of the lettuce plant according to the flowering characteristics thereof. For example, since mayonnaise and corydalis impatiens have continuous flowering characteristics in which they bloom about 1.5 months after sowing in spring and then continue to open to late autumn, it is not necessary to match the flowering period of the lettuce plant, but the procedure can be made efficient by sowing and planting the lettuce plant at the same time. Therefore, it is desired that the Melanhua and the corydalis impatiens be sown and planted simultaneously with the lettuce plants. On the other hand, when a plant species having low continuous flowering performance is used as the auxiliary honey source plant, it is necessary to appropriately adjust the planting period so as to match the flowering period of the lettuce plant in accordance with the flowering characteristics.

The ratio of the number of established varieties of the auxiliary honey-source plants to the lettuce plants is not particularly limited, and can be arbitrarily adjusted depending on the type of the auxiliary honey-source plants and the cultivation environment. For example, the ratio of the number of colonizations is lettuce: the auxiliary honey source plant ranges from 2:1 to 20:1, and is preferably a lettuce plant: the auxiliary honey source plant is 2: 1-4: 1, and more preferably a lettuce plant: the auxiliary honey source plant is about 5: 2.

The site for permanent planting of the auxiliary honey plant is desirably the periphery of the nest of the bees as a pollen propagation means, but may be disposed dispersedly in or around the nursery.

Active floral-interview activities of wasps of the genus aphelesia (e.g., alfalfa aphelenchoides) typically require temperatures above 25 ℃. Therefore, it is desired to cultivate lettuce plants for seed collection in an environment where the temperature during the flowering period is 25 ℃ or higher diurnally.

Because of the homing properties of the bees of the genus Ceratoptera (e.g., alfalfa cutting bees), not only closed net rooms but also open gardens and the like can be utilized. That is, the present invention may be implemented in any one of an open or closed nursery facility. Therefore, if lettuce plants that may cross are not present in the surroundings, it is desirable to use the lettuce plants in an open nursery that is inexpensive to produce.

Methods for incubating and stocking bees belonging to the genus Apis can be routinely employed by those skilled in the art. For example, reference 3 and the like can be referred to as appropriate. The method for incubating and stocking the alfalfa leaf cutting bees can be performed according to a conventional method for harvesting alfalfa.

When bees of the genus Cerebrophora (for example, lucerne Cerebrophora) are used for the breeding of the genus Lactuca, stocking is preferably performed at intervals of 1 to 2 weeks.

In addition, regarding the desired stocking density of the bees of the genus serenoa (e.g., alfalfa serenoa), 5 or more per 1 st is desired for each stocking, but it is also influenced by the line difference of the lettuce plant used and the cultivation environment, and therefore it is preferable to appropriately adjust the stocking density in consideration of the cost.

Stocking of bees of the genus aphelesia (e.g., alfalfa aphelenchoides) is preferably performed in the evening of the previous day or in the early morning before sunrise. Since the pollen parent of the lettuce plant stops flowering immediately after setting, it is desirable to appropriately cut the pollen parent so that the pollen parent can continuously flower and supply pollen for a long period of time.

[ reference documents ]

[ reference 1] W.P.KEMP et al, "Development and expression of the Alfa Polifier Mega project", "antibiotics of the environmental Society of America, 93(4):904-

[ reference 2] Eric Mader et al, "manufacturing Alternative Poliners: A Handbook for Beekeepers, Growers, and Conservationists", SARE Handbook 11(2010), p10

[ reference 3] Colophonium photo male, "utilization of insect utilization science series 7 pollinators", 1 st edition, science house Co., Ltd., 2.1996, p.97-112

[ reference 4] Dave Goulson, "EFFECTS OF INTRODUCED BEES ON NATIVEECOSYSTEMS", Annu.Rev.Ecol.Evol.Syst.34:1-26(2003), p6,

[ reference 5] "American Musum of Natural History, Bee specific Recorddatabase", ID _ parent AMNH _ BEE00026543, [ on-line ], American Musum of NaturalHistory, [ 11/6 th-day search in 2017 ], Internet (http:// www.discoverlife.org/mp/20l

[ reference 6] "American Musum of Natural History, Bee specific Recorddatabase", ID _ parent AMNH _ BEE00015546 [ on line ], American Musum of NaturalHistory [ search on 6/11/2017 ], Internet (http:// www.discoverlife.org/mp/20l

[ reference 7] John Pickering, "DISCOVER LIFE", [ on-line ], [ 11/6 th-day search in 2017 ], Internet (http:// www.discoverlife.org/mp/20q)

[ reference 8] "American Musum of Natural History, Bee specific Recorddatabase", ID _ parent AMNH _ BEE00220721, [ on-line ], American Musum of NaturalHistory, [ 11/6 th-day search in 2017 ], Internet (http:// www.discoverlife.org/mp/20l

[ reference 9] "Bee Biology and Systematics Laboratory databa", ID _ parent BBSL200815, [ on-line ], Bee Biology and Systematics Laboratory, [ 11/6/11/2017 ], Internet (http:// www.disco verlife. org/mp/20

[ reference 10] "American Musum of Natural History, Bee Sp environmental Recorddatabase", ID _ parent AMNH _ BEE00107046, [ on-line ], American Musum of NaturalHistory, [ 11/6-th-2017 ] Internet (http:// www.discoverlife.org/mp/20?id ═ AMNH _ BEE 00107046; A MNH _ BEE00107116)