JP3386292B2 - Method for producing F1 tomato plant with restored fertility - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for producing an F1 tomato plant having fertility by crossing a tomato plant having a cytoplasmic male sterility gene with a tomato plant having a fertility recovery gene, and a cytoplasm. The present invention relates to a fertile tomato plant having a male sterility gene and a fertility recovery gene.

[0002]

2. Description of the Related Art At present, in the production of F1 seeds for vegetables, the cabbage (Brassica ole) belonging to the Cruciferae family is used.
racea), Japanese radish (Raphanus sativus) and Chinese cabbage (Br
The property of self-incompatibility is utilized, such as assica pekinensis). Also, carrots (Daucus carota) belonging to the Umbelliferae family and leeks (Allium fistulosum) belonging to the Liliaceae family (Lumaceae) produce cytoplasmic male sterility to produce F1 seeds. Such self-incompatibility and cytoplasmic male sterility are genetically controlled, and are important techniques for F1 seeding of horticultural crops.

Regarding the F1 seeding of tomatoes, there are very few cases in which the above-mentioned self-incompatibility and cytoplasmic male sterility are utilized, and in order to produce F1 seeds, the anthers of the female line for seed production are removed before flowering ( After the flowering, the pollen of the male line was crossed. Therefore, a great deal of labor was required for F1 seed production. In order to solve this problem, the present inventors have created a tomato having cytoplasmic male sterility by asymmetric cell fusion of cultivated tomato and Solanum acaule Hawkes of Solanum plant. Was filed (Japanese Patent Application Laid-Open No. 2-138927).
Issue). However, since the above-mentioned tomato is male-sterile, seeds cannot be formed by selfing. In the case of tomatoes, seeds play an important role in fruit growth, and in the absence of seeds, it is not possible to obtain full fruit. Therefore, it has been desired to develop a means for recovering the fertility of F1 seeds.

[0004]

SUMMARY OF THE INVENTION An object of the present invention is to provide means for producing an F1 tomato plant having both a cytoplasmic male sterility gene and a fertility recovery gene.

[0005]

Means for Solving the Problems As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that some strains of wild species related to tomato and cultivated tomato have a fertility recovery gene. And completed the present invention. That is, the present invention provides a tomato plant having a cytoplasmic male sterility gene as a seed parent and a tomato plant having a fertility recovery gene as a pollen parent, and mating them to produce an F1 tomato plant having fertility. It is a method for producing an F1 tomato plant having characteristic fertility.

Further, the present invention is a tomato plant having fertility, having a cytoplasmic male sterility gene and a fertility recovery gene. Hereinafter, the present invention will be described in detail. The tomato plants of the present invention are not limited to cultivated tomatoes (Lycopersicon esculentum), and include plants that broadly belong to the genus Lycopersicon. The tomato plants of the present invention also include plants obtained by mating lycoperisicon plants with other plants, for example, Solanum plants, cell fusion, and the like.

In the present invention, the fertility restoring gene refers to a gene that restores fertility of a male-sterile tomato plant. Tomato plants with this gene include Lycopersicillin pinpinerifolium (L
ycopersicon pimpinellifolium) ・ LA1670 strain, Lycopersicon esculentum cerasiforme (Lycopersi
con esculentum var.cerasiforme) LA1673 strain, Lycopersicon cheesmanii
) ・ LA166 strain, cultivar Lycopersicon esculentum (Lycopersicon esculentum) ・ MN1-5 strain (hereinafter, for these, "LA167
It is abbreviated as "0", "LA1673", "LA166", and "MN1-5". ). LA1670, LA1673 and LA166 are TOMATO GEN
ETICS STOCK CENTER, DEPARTMENT OF VEGETABLE CROPS,
UNIVERSITY OF CALIFORNIA, DAVIS, CALIFORNIA 9561
6, MN1-5 was obtained from Tokita, and seeds of any strains are currently owned by the applicant and may be distributed as necessary. it can.

In the present invention, the cytoplasmic male sterility gene is a gene contained in the cytoplasm and expresses a male sterility trait. Examples of plants containing this gene include the MSA1 line (hereinafter,
Abbreviated as "MSA1". ). MSA1 was obtained from Nichirei Co., Ltd. The seeds are currently owned by the applicant and can be distributed if necessary.

The tomato plant having fertility of the present invention can be obtained by crossing the tomato plant having the cytoplasmic male sterility gene as a seed parent and the tomato plant having the fertility restoring gene as a pollen parent. The tomato plant having fertility of the present invention has no apparent difference from the tomato plant having normal fertility in appearance, but it has normal fertility in that it has both a fertility recovery gene and a cytoplasmic male sterile gene. Is distinctly different from tomato plants with. That is, a normal tomato plant does not develop an individual having a male sterility trait no matter how many times it repeats self-fertilization. Individuals separate at a specific ratio. In addition, LA
Seeds obtained by crossing 1670 with MSA1, LA1673 and MSA1
Seeds obtained by crossing with, seeds obtained by crossing with LA166 and MSA1, seeds obtained by crossing with MN1-5 and MSA1 are currently possessed by the applicant, and if necessary. Can be sold.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in more detail with reference to Examples. [Example 1] MSA1 which is a cytoplasmic male sterile line was pollinated with pollen of a wild strain of 15 wild species related to tomato, and the fertility of the obtained F1 plant was examined. That is, F1 plants were selfed and the number of fruits and seeds obtained was examined, and the average number of seeds per fruit was determined from them. The results are shown in Table 1. In addition,
All closely related wild species used for mating had different collection sites, and even within the same species, a clear difference in shape could be confirmed.

[0011]                 Table 1 Detection of fertility recovery genes for MSA1 ──────────────────────────────────── Male strain name Species name Collection site Number of fruits Number of seeds 1 Average number of seeds ──────────────────────────────────── LA 166 L. cheesmanii Ecuador 13 225 17 LA 716 L. pennellii Peru 14 0 0 LA 722 L. pimpinelifolium Peru 11 4 0.4 LA1223 L. hirsutum Ecuador 11 1 0.1 LA1306 L. chmielewskii Peru 5 0 0 LA1326 L.parviflorum Peru 7 0 0 LA1670 L. pimpinellifolium Peru 22 738 34 LA1673 L.esc.var.cerasiforme Peru 19 331 21 LA1926 L. pennellii Peru 9 0 0 LA2097 L. pimpinellifolium Ecuador 19 2 0.1 LA2194 L.parviflorum Peru 9 0 0 LA2657 L. pennellii Peru 20 2 0.1 LA2666 L.esc.var.cerasiforme Peru 300 LA2695 L. chmielewskii Peru 7 0 0 LA2791 L.esc.var.cerasiforme Colombia 0 0 0 MN1-5 L. esculentum Tokita 14 51 4 ────────────────────────────────────

The number of fruits and the number of seeds represent the total number obtained by selfing, and the average number of seeds per fruit represents the average number of seeds per fruit obtained by the number of seeds ÷ the number of fruits. As shown in Table 1, F1 plants obtained by mating LA1670, LA1673 and LA166 as male lines had completely recovered fertility. From this, it is considered that LA1670, LA1673 and LA166 have fertility restoring genes.

[Example 2] Cytoplasmic male sterile line OF209CMS
-BC and MNY-1CMS-BC to pollinate pollen of 2 wild strains of tomato closely related fertility recovery gene (LA1670, LA1673) and 1 line of cultivated tomato having fertility recovery gene (MN1-5) The fertility of the obtained F1 plant was examined. That is, F1 plants were selfed and the number of fruits and seeds obtained was examined, and the average number of seeds per fruit was determined from them. The results are shown in Table 2. The OF209CMS-BC was backcrossed to obtain OF209 (Tokita's variety "Oomiya 16
3 ”female parental line), which is a male-sterile cytoplasmic line of MSA1 introduced into the male sterile cytoplasm, and MNY-1CMS-BC becomes MNY-1 (female parental line of Tokita's variety“ Sungold ”). It is a male-sterile cytoplasmic line introduced with the male sterile cytoplasm of MSA1.

Table 2 Recovery of fertility against OF209CMS-BC and MNY-1CMS-BC ──────────────────────────────── Mating combination Average number of seeds per fruit ──────────────────────────────── OF209CMS-BC × MN1-5 4.8 OF209CMS-BC x LA1670 11.4 OF209CMS-BC x LA1673 31.6 MNY-1CMS-BC x MN1-5 0.1 MNY-1CMS-BC x LA1670 7.6 MNY-1CMS-BC x LA1673 18.8 ── ────────────────────────────── As shown in Table 2, LA1670, LA1673 and MN1-5 were crossed as male lines. The F1 plant thus obtained had completely recovered fertility.

[Example 3] Hybrid F2 between MSA1 and LA1670
We investigated the phenotypic segregation ratio for fertility recovery in the population. The results are shown in Table 3. Table 3 F2 analysis of MSA1 × LA1670 ───────────────────────────────── Fertile population Sterile population Kai Square (3: 1) P (df = 1) ───────────────────────────────── 65 13 2. 889. 05 <P <. 1 ───────────────────────────────── As shown in Table 3, the separation of fertile and sterile individuals Ratio is 3:
Therefore, it can be presumed that the fertility recovery gene is controlled by one dominant gene. Further, it can be presumed that this mode of fertility recovery is sporophyte-type fertility recovery.

[Example 4] 32 lines possessed by Tokita were crossed with MSA1 and the recovery of fertility of the obtained F1 plant was investigated. As a result, one line of cherry tomato (L. esculentum), MN1- 5 showed fertility recovery to MSA1 (Table 1). Further, OF209C obtained by cytoplasmic replacement in the same manner as in Example 2
When fertility recovery for MS-BC and MNY-1CMS-BC was examined, fertility recovery was observed as in the case of closely related wild species (Table 2).

Therefore, the sympatricity of the fertility recovery genes of this cultivated tomato MN1-5 and the wild-type related species was tested. MN1-
5 and LA1670, MN1-5 and LA1673 are crossed and F1
I got a plant. The pollen of both F1 plants was pollinated by MSA1 and the fertility of the resulting progeny plant was examined. All of the plants derived from LA1670 show recovery of fertility, whereas LA1670
From 673, plants in which fertility was not recovered were isolated. This indicates that the fertility restoration gene carried by MN1-5 exists on the same chromosome as the fertility restoration gene carried by LA1670, and the fertility restoration gene carried by LA1673 exists on a different chromosome. Sedentary).

[Example 5] Two male sterile lines having a male sterile cytoplasm different from MSA1 (OF209CMS-F, PSFM-CMS-F)
F was obtained by pollinating the pollen of two tomato wild-type wild strains (LA1670, LA1673) having a fertility recovery gene and one cultivated tomato line (MN1-5) having a fertility recovery gene.
The fertility of one plant was investigated. That is, F1 plants were selfed and the number of fruits and seeds obtained was examined, and the average number of seeds per fruit was determined from them. The results are shown in Table 4.

OF209CMS-F is a cytoplasmic male sterile line produced by asymmetric cell fusion of OF209 (Tokita's variety, "Omiya 163" female parent line) and Solanum acaule Hawkes. And PSFM-CMS
-F is a cytoplasmic male sterile line created by asymmetrical cell fusion of Tokita's conserved line PSFM and Solanum Akaure Hawks.

[0020]       Table 4 Recovery of fertility against cytoplasmic male sterile lines other than MSA1 ───────────────────────────────       Mating combination Average number of seeds per fruit ───────────────────────────────   OF209CMS-F × MN1-50 (sterile)   OF209CMS-F × LA1670 0 (sterile)   OF209CMS-F × LA1673 0 (sterile)   PSFM-CMS-F x MN1-50 (sterile)   PSFM-CMS-F x LA1670 8   PSFM-CMS-F x LA1673 28 ────────────────────────────────

As shown in Table 4, in the case of OF209CMS-F,
No fertility is recovered when crossed with any of the lines, whereas in the case of PSFM-CMS-F, when crossed with MN1-5,
There is no recovery of fertility, but when crossed with LA1670 and LA1673, recovery of fertility is seen as in the case of MSA1.
Changes in fertility recovery were observed due to different cytoplasmic composition. In asymmetric cell fusion, the cytoplasmic composition of the obtained plant body (in many cases, structural changes in the mitochondrial genome) is known, and therefore fertility may not be restored due to the difference in mitochondrial DNA recombination. I think there is. However, since there is also a male sterile cytoplasm in which fertility is restored, the usefulness of the fertility restoration genes possessed by LA1670 and LA1673 was shown.

Further, although it was described in Example 4 that the fertility recovery genes of MN1-5 and LA1670 are present on the same chromosome, the fertility recovery properties for the cytoplasmic male sterile line PSFM-CMS-F are different. Therefore, it was shown that the fertility recovery genes possessed by MN1-5 and LA1670 are not the same.

[Example 6] Since the restoration of fertility is dominated by one dominant gene, by introducing the fertility restoration gene into a strain that does not carry the fertility restoration gene,
A fertility recovery line can be easily created. Therefore, we tried to introduce the fertility recovery gene possessed by the cultivar MN1-5 into the cultivar June Pink as follows.

First, an F1 plant obtained by crossing June Pink (seed parent) and MN1-5 (pollen parent) was backcrossed with June Pink to obtain a first backcross generation (BC1). Then
In order to confirm the presence or absence of the fertility recovery gene, this backcross first generation (pollen parent) was crossed with MSA1 (seed parent), and the obtained individuals (individual numbers 1 to 13) were selfed and The average number of seeds per fruit was calculated for each individual. The results are shown in Table 5.

[0025]                         Table 5 ────────────────────────────   Individual number Fruit set Number of seeds Average number of seeds per fruit ────────────────────────────       1 4 4 1.0       2 0 0       3 0 0       4 50       5 40       6 9 2 0.2       7 9 1 0.1       8 30       9 3 1 0.3     100     11 90     12 20     13 90 ────────────────────────────

As shown in Table 5, seeds were formed in individuals Nos. 1, 6, 7 and 9 but not in other individuals. That is, the fertility recovery gene derived from MN1-5 was introduced into only the above 4 individuals. For these 4 individuals,
Backcrossed with June Pink. The second-generation traits of the obtained backcrosses were changed to the June Pink type, indicating that it is possible to produce June Pink into which the fertility restoration gene has been introduced.

In the same manner as above, Tokita storage system PS
The first generation of backcrosses of M with LA1670 and LA1673 was obtained.
This was crossed with a cytoplasmic male sterile line PSFM-CMS-F, and the obtained individuals were selfed and the average number of seeds per fruit of all the individuals was determined. The results are shown in Table 6.

[0028]                                 Table 6 ───────────────────────────────────       Mating combination 1 Average number of seeds ───────────────────────────────────  First generation of backcross between PSFM-CMS-F x PSM and LA1670 63  First generation backcross between PSFM-CMS-F x PSM and LA1673 24 ───────────────────────────────────

[Reference Example 1] Production amount of F1 seeds by normal emasculation and F1 using a cytoplasmic male sterile line
The seed production was compared. That is, OF209CMS-BC, which is a cytoplasmic male sterile line, was pollinated with pollen of cultivated tomato Z102-N line, the numbers of fruits and seeds obtained were examined, and the average seed amount per fruit was determined from them. Further, the fertile line OF209 was emasculated before anthesis before anthesis, then pollinated with the pollen of the cultivated tomato Z102-N line, and the average seed amount per fruit was determined in the same manner as described above. The results are shown in Table 7.

[0030]                                 Table 7 ────────────────────────────────────     Mating combination Individual number Fruit number Seed amount Average seed amount per fruit ────────────────────────────────────  OF209CMS-BC x Z102-N 5 35 44 ml 1.3 ml  OF209 (excluding male) × Z102-N 2 7 6.5 0.9 ────────────────────────────────────

As shown in Table 7, when the cytoplasmic male sterility was used, the production amount of F1 seeds was increased as compared with the usual seeding method. It was speculated that this is because the stress caused by emasculation was reduced, and the effectiveness of utilizing cytoplasmic male sterility was shown.

[0032]

INDUSTRIAL APPLICABILITY The present invention provides means for producing F1 tomato plants having both a cytoplasmic male sterility gene and a fertility recovery gene.

─────────────────────────────────────────────────── ─── Continuation of the front page (56) References JP-A-2-138927 (JP, A) JP-A-6-501613 (JP, A) JP-A-3-503004 (JP, A) (58) Field (Int.Cl. ⁷ , DB name) A01H 1/00 A01H 5/00 C12N 5/10

Claims (6)

(57) [Claims] 1. A method for producing an F1 tomato plant having fertility, which comprises using as a seed parent a cytoplasmic male sterile line introduced by backcrossing MSA1 or MSA1 male sterile cytoplasm. b)
The method as described above, characterized in that the above are crossed as pollen parents. (a) LA1670, LA1673, LA166, or MN1-5 (b) The tomato plant was backcrossed to an F1 plant obtained by crossing the tomato plant with (a) as a pollen parent to introduce a fertility restoration gene. Fertility recovery line 2. A method for producing an F1 tomato plant having fertility, which is characterized in that PSFM-CMS-F is used as a seed parent and (a) or (b) below is used as a pollen parent. The above method. (a) LA1670 or LA1673 (b) A fertility recovery line in which a fertility recovery gene is introduced by backcrossing the tomato plant to an F1 plant obtained by crossing the tomato plant with (a) as a pollen parent. 3. A cytoplasmic male sterile line obtained by backcrossing the male sterile cytoplasm of MSA1 into OF209CMS-BC or MNY-1.
The method of claim 1, which is CMS-BC. 4. A method for producing an F1 tomato plant having fertility, which is characterized by crossing with MSA1 as a seed parent and the following (a) or (b) as a pollen parent. (a) LA1670, LA1673, LA166, or MN1-5 (b) June Pink was used as a seed parent and MN1-5 was used as a pollen parent to obtain a fertility recovery gene by backcrossing June Pink to an F1 plant. Fertility recovery line 5. A method for producing an F1 tomato plant having fertility, which comprises mating OF209CMS-BC or MNY-1CMS-BC as a seed parent and LA1670, LA1673 or MN1-5 as a pollen parent. 6. A method for producing a F1 tomato plant having fertility, which is characterized in that PSFM-CMS-F is used as a seed parent and the following (a) or (b) is used as a pollen parent. The above method. (a) LA1670 or LA1673 (b) A fertility recovery line in which a fertility recovery gene was introduced by backcrossing PSM to an F1 plant obtained by crossing PSM with (a) above as a pollen parent