Literature DB >> 33603559

Development of 'Darumadattan', a semidwarf lodging-resistant Tartary buckwheat cultivar, using gamma-ray irradiation.

Akemi Shimizu1, Hiroyasu Yamaguchi1,2, Konosuke Degi1,3, Toshikazu Morishita1.   

Abstract

We developed a new cultivar of Tartary buckwheat (Fagopyrum tataricum Gaertn.), 'Darumadattan'. This is the first semidwarf Tartary buckwheat cultivar to be developed by mutation breeding using gamma-ray irradiation. In 1999, 100 dry seeds of the leading Japanese cultivar, 'Hokkai T8' (known at that time as 'Hokkei 1'), were gamma-ray-irradiated with a total dose of 500 Gy (25 Gy/h × 20 h) at the Institute of Radiation Breeding (IRB), National Institute of Agrobiological Sciences, Hitachiomiya, Ibaraki, Japan. The seeds were sown in August 1999 in a field at IRB, and M2 seeds were collected from the eight individual plants that survived. In August 2000, 240 M2 seeds were sown in a field, and one semidwarf plant was found. The line named 'IRBFT-20' developed from the selected plant was investigated for its semidwarf characteristic and genetic stability in 2001-2005. 'IRBFT-20' was submitted for registration in 2011 and registered as the cultivar 'Darumadattan' in 2013. This name was chosen because the plants resemble "Daruma dolls" and "dattan" means "Tartary" in Japanese. 'Darumadattan' is a highly lodging-resistant and high-yielding cultivar and is expected to be used as both a commercial cultivar and a crossing parent.
Copyright © 2020 by JAPANESE SOCIETY OF BREEDING.

Entities:  

Keywords:  Darumadattan; Tartary buckwheat; gamma-ray irradiation; lodging resistance; mutant; semidwarf

Year:  2020        PMID: 33603559      PMCID: PMC7878941          DOI: 10.1270/jsbbs.20044

Source DB:  PubMed          Journal:  Breed Sci        ISSN: 1344-7610            Impact factor:   2.086


Introduction

Since it contains large amounts of protein and minerals, buckwheat is well known as a healthy food. Specifically, the rutin content in Tartary buckwheat seeds is approximately 100 times that in the seeds of common buckwheat (Fagopyrum esculentum Moench) (Morishita ). Although buckwheat is consumed in Japan, Tartary buckwheat is unpopular because of its bitter taste (Kawakami ). The bitterness of Tartary buckwheat derives from the presence of three bitter compounds, quercetin and uncertain F3 and F4 compounds (Kawakami ). Quercetin, a bitter compound, is produced from the degradation of rutin by rutinosidase which catalyze rutin to quercetin and rutinose (Yasuda ). However, the trace-rutinosidase cultivar ‘Manten-Kirari’ has overcome the defects of bitterness in Tartary buckwheat (Suzuki ), enabling the production of rutin-rich and high antioxidative activity foods (Ishiguro , Suzuki , 2019). Furthermore, the cultivar is promising as a good rutin resource (Morishita ). Rutin is commonly known as a functional component in Tartary buckwheat and has biological functions, such as antihypertension activity (Matsubara ); the reduction of serum myeloperoxidase and cholesterol levels (Wieslander ), mucosal symptoms, headaches, and tiredness (Wieslander ); and the reduction of body weight, BMI (body mass index), and TBARS (2-thiobarbituric acid reactive substances) levels (Nishimura ). Because of these favorable traits, Tartary buckwheat has attracted attention in Japan and the breeding objective of improving yielding ability was discussed at a meeting of the Crop Science Society of Japan (Michiyama ). Yet only a few Tartary buckwheat cultivars are available: 13 cultivars were registered in Japan from 2006 through 2020, and one cultivar has been under examination in May 2020 (Ministry of Agriculture, Forestry and Fisheries 2020). The cultivar ‘Hokkai T8’, the first major cultivar in Japan, showed high plant height and easy lodging (Honda ). Later, the cultivar ‘Hokuriku 4’, which is 10 cm shorter than ‘Hokkai T8’, was developed (Ito ). The plant height of ‘Manten-Kirari’ was similar to that of ‘Hokkai T8’ (Suzuki ). As these available Tartary buckwheat cultivars are tall and lodge easily, lodging resistance is an important objective in Tartary buckwheat breeding (Hagiwara , Inoue ). Semidwarf traits have been promising material for lodging resistance in common buckwheat (Morishita ). As common and Tartary buckwheat have a similar plant shape, the semidwarf trait is expected to have the same effect in Tartary buckwheat. Dwarf and semidwarf genes have been reported in common buckwheat (Minami , Morishita , Ohnishi and Nagakubo 1982), but the practical semidwarf cultivars have not been developed. Available semidwarf trait have never been discovered in the genetic resources of Tartary buckwheat. However, semidwarf materials were developed by mutation breeding, and two semidwarf genes, sdA and sdB have been identified in Tartary buckwheat (Morishita ). We developed a semidwarf cultivar ‘Darumadattan’ which belongs to the sdA group. ‘Darumadattan’ was registered in 2013 (Ministry of Agriculture, Forestry and Fisheries 2013a). Here, we report on the breeding procedures and characteristics of ‘Darumadattan’.

Materials and Methods

Plant materials

Tartary buckwheat cultivar ‘Hokkai T8’ was used as the original cultivar. In July 1999, 100 dry seeds of ‘Hokkai T8’, known at that time as ‘Hokkei 1’ (before its registration as ‘Hokkai T8’), were acutely gamma-ray-irradiated with a total dose of 500 Gy (25 Gy/h × 20 h) in the gamma room of the Institute of Radiation Breeding (IRB) at the National Institute of Agrobiological Sciences (Existing NARO Institute of Crop Science, Radiation Breeding Division), Hitachiomiya, Ibaraki, Japan.

Breeding

The procedure is summarized in Table 1. All cultivation was carried out in the experimental field at IRB (Kamimurata, Hitachiomiya, Ibaraki, Japan; 140°24ʹE, 36°29ʹN). The cultivation conditions are shown in Table 2. All irradiated M1 seeds were sown in August 1999. At harvest time in November 1999, eight plants of the M1 generation had survived, and M2 seeds were collected from them individually. Thirty M2 seeds from each M1 plant were sown as individual lines in August 2000. One semidwarf M2 plant was found and named ‘R-21-8’. Its progeny was separated into two distinguishable lines (with or without red leaves) during fixation and confirmation at the M3 generation in 2001. These lines were named ‘R-21-8-1’ and ‘R-21-8-2’ in the M4 generation. However, as these two lines showed no difference during cultivation in 2002, it seemed that the difference observed in 2001 was caused by environmental factors. As ‘R-21-8-1’ yielded no seeds, ‘R-21-8-2’ was selected in 2003 at M6. At the M7 generation, grown in 2004, the selected line was named ‘IRBFT-20’, and seeds of ‘IRBFT-20’ were propagated for subsequent performance test from 2004 to 2006.
Table 1.

Breeding process of cultivar ‘Darumadattan’

Year19992000200120022003200420052006200720082013
Breeding procedureAcute 500 Gy irradiation for dry seedsPropagationIndividual selectionFixation and confirmationFixation and confirmationLine selectionPropagation of seedsPerformance test and propagation of seedsPerformance testRegistration
Number of materials100 seeds (M0)100 seeds8 lines, 30 seeds/line1 line2 lines2 lines1 line1 line1 line1 line
Number of selections100 seeds (M1)8 plants survived1 plant2 separated lines2 lines1 line
GenerationM0 and M1M2M3M4M5M6M7 and M8
NameR-21-8R-21-8-1R-21-8-2R-21-8-1R-21-8-2R-21-8-2IRBFT-20IRBFT-20IRBFT-20Darumadattan
Table 2.

Cultivation conditions in Ibaraki (IRB)

PurposeYearNumber of replicationsArea (m2)Interrow space (cm)Seeding density (seeds/m2)Fertilizer (kg/a)
NP2O5K2O
Selection1999 to 2005160 260.260.260.26
Performance test2006 to 200824.860 260.280.280.28
Seeding density test200624.8303030606060 52104208 26 521040.280.280.28
Heavy fertilizer application test200624.86030 26 521.401.401.40

Performance tests at the IRB in Ibaraki

Performance tests for agronomic characteristics were carried out in the experimental field at IRB in 2006–2008. Cultivation conditions for the tests are shown in Table 2. Fertilizer containing N, P2O5 and K2O at 0.26, 0.26, and 0.26 kg/a, respectively, was applied in 1999–2005 and with 0.28, 0.28, and 0.28 kg/a, respectively, was applied in 2006–2008. Characteristics were evaluated according to the Test Guidelines of Buckwheat (Ministry of Agriculture, Forestry and Fisheries 2013b). Degree of lodging was determined by observation; no lodging was 0 and completely lodged (lying completely flat) was 5. Values 1 to 4 were divided equally between 0 and 5.

Seeding density and heavy fertilizer application tests at the IRB in Ibaraki

Seeding density and heavy fertilizer application tests were carried out in 2006. Cultivation conditions for the tests are shown in Table 2. Standard fertilizer containing N, P2O5 and K2O at 0.28, 0.28, and 0.28 kg/a, respectively, was applied and heavy fertilizer, 5 times the standard, containing 1.40, 1.40, and 1.40 kg/a, respectively, was applied. Characteristics were evaluated according to performance tests.

Performance tests in Hokkaido

In 2009, a performance test was carried out in the experimental field at Memuro Upland Farming Research Division, Hokkaido Agricultural Research Center (HARC), NARO (Shinsei, Memuro, Kasai-gun, Hokkaido, Japan; 143°03ʹE, 42°55ʹN). Seeds were sown on June 4 under the following conditions: plot size, 2.4 m2; seeding density, 66.7 seeds/m2; row spacing, 60 cm; 2 replications. Fertilizer containing N, P2O5, K2O and MgO at 0.18 kg/a, 0.72 kg/a, 0.42 kg/a and 0.15 kg/a, respectively was applied. Agronomic characteristics were evaluated according to the Test Guidelines of Buckwheat (Ministry of Agriculture, Forestry and Fisheries 2013b).

Performance tests in Nagano

In 2014, a performance test was carried out in the experimental field at the Nagano Vegetable and Ornamental Crops Experimental Station (Souga, Shiojiri, Nagano, Japan; 137°56ʹE, 36°6ʹN). Seeds were sown on August 4 under the following conditions: plot size, 7.2 m2; seeding density, 150 seeds/m2; row spacing, 30 cm; 2 replications. Characteristics were evaluated according to the guidelines of Nagano Prefecture (Nagano Prefecture 2010).

Rutin analysis

Seeds of plants from the sowing density and heavy fertilizer application tests were used. The rutin content of ‘IRBFT-20’ and ‘Hokkai T8’ seeds were determined using the method of Morishita .

Analysis of variance (ANOVA)

ANOVA was performed using the programs Excel (Microsoft Co., USA) and Excel statistics (Social Survey Research Information Co., LTD., Japan).

Wind data

Data for wind velocity was obtained from the website of the Japan Meteorological Agency (Japan Meteorological Agency 2020). Before 2007, maximum instantaneous wind velocity was not published. The data for Ibaraki was obtained from Hitachiomiya in 2008. The data of Nagano was obtained from Matsumoto Imai, near Shiojiri, in 2014. The data for Hokkaido was obtained from the observed values at Memuro Upland Farming Research Division, HARC, NARO.

Application for registration

‘IRBFT-20’ was submitted for registration in 2011 under the name ‘Darumadattan’. In 2013, ‘Darumadattan’ was officially registered as a cultivar of Tartary buckwheat with the Ministry of Agriculture, Forestry and Fisheries (Ministry of Agriculture, Forestry and Fisheries 2013a). The name ‘Darumadattan’ was derived from “Daruma doll”, which the plants resemble, and “dattan”, meaning “Tartary” in Japanese.

Results

Performance tests

The results of performance tests, ANOVA and wind velocity during performance tests are shown in Tables 3, 4 and in Fig. 1, respectively. In Ibaraki, before maturing, there were a few days with strong wind before maturity in 2008. ‘Darumadattan’ plants were shorter than ‘Hokkai T8’ by approximately 50 cm, and their growing period was longer by 6 days in 2006 and 2008, and 4 days in 2007. The number of nodes on the main stem of ‘Darumadattan’ exceeded that of ‘Hokkai T8’ but no significant difference was observed. The difference in seed yield between ‘Darumadattan’ and ‘Hokkai T8’ was not significant and varied with the cultivation year. Both ‘Darumadattan’ and ‘HokkaiT8’ can achieve high yield in Ibaraki. The 1000 seed weight was significantly higher in ‘Hokkai T8’ than in ‘Darumadattan’.
Table 3.

Results of performance test

PlaceYearSowing time month.dayFlowering time month.dayTime of maturity month.dayPlant height cmNumber of nodes on main stem No./plantNumber of primary branches No./plantDegree of lodgingaStem diameter mmSeed yield kg/a1000 seed weight g
DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8
Ibaraki20068.189.249.2211.1011.046013822.220.77.37.00317.617.715.219.0
20078.2810.0510.0111.2411.205311520.118.66.96.413.818.116.621.4
20088.209.269.2211.2011.14447317.515.57.06.0037.89.515.520.6
Average8.229.289.2511.1811.135210919.918.37.16.50313.115.115.820.3
Hokkaido20096.047.197.179.109.096717920.219.05.94.6048.56.919.336.913.620.0
Nagano20148.049.119.0910.29b10.29b5615316.917.65.14.40328.819.814.919.7

There was no replication of ‘Darumadattan’ in 2007 due to moisture damage.

–: No data.

0 none–5 completely lodgied.

Damage by frost on October 29 prevented further maturation.

Table 4.

ANOVA of performance test

PlaceFactorPlant heightNumber of nodes on main stemNumber of primary branchesSeed yieldStem diameter1000 seed weight
IbarakiaCultivar (C)**NSNSNS**
Year (Y)**NSNSNS*
C × Y*NSNSNSNS
HokkaidoCultivar**NSNS*****
NaganoCultivar**NSNS***

Analysis of 2006 and 2008.

*, **, NS: significantly different at the 0.05 and 0.01 levels and non-significant at the 0.05 level.

Fig. 1.

The wind velocity during cultivation periods. The wind velocity of places during cultivation period. Wind velocity data of Ibaraki and Nagano were obtained from the web site of Japan Meteorological Agency (JMA) and that of Hokkaido was obtained from the observed values at Memuro Upland Farming Research Division HARC NARO. S: seeding, F: flowering, M: maturity.

In Hokkaido, calm days prevailed during cultivation. ‘Darumadattan’ plants were shorter than ‘Hokkai T8’ by approximately 110 cm, and their main stems were significantly thicker. The degree of lodging of ‘Darumadattan’ was very low. The seed yield and 1000 seed weight were higher in ‘Hokkai T8’ than in ‘Darumadattan’. Other characteristics were similar in both cultivars. In Nagano, there were many days with strong wind during the growing period and two typhoons hit during the maturing period. ‘Darumadattan’ plants were shorter than ‘Hokkai T8’ by approximately 100 cm. Their time of flowering was similar, but both cultivars were damaged before maturity by frost on October 29, making it impossible to determine the time of maturity. The seed yield of ‘Darumadattan’ was obviously higher than that of ‘Hokkai T8’. The 1000 seed weight was higher in ‘Hokkai T8’ than in ‘Darumadattan’. ‘Hokkai T8’ lodged in strong wind, but ‘Darumadattan’ did not lodge.

Seeding density and heavy fertilizer application tests

The results of seeding density and heavy fertilizer application tests are shown in Table 5. In seeding density tests, ‘Darumadattan’ plants were approximately 50% shorter than ‘Hokkai T8’ plants, regardless of the interrow space or seeding density. The degree of lodging of ‘Darumadattan’ was very low, regardless of the interrow space or seeding density. Although no significant difference in seed yield was observed among cultivation treatments, the following differences were observed. At the same seeding density, reducing the interrow space from 60 cm to 30 cm caused an increase in the yield of ‘Darumadattan’ but not in ‘Hokkai T8’. At a given row spacing, increasing the seeding density did not increase yield in either cultivar. The rutin content of ‘Darumadattan’ was significantly lower than that of ‘Hokkai T8’ in all treatment. On the other hand, the differences of the rutin content between treatment were not significant. Even when the amount of fertilizer was increased (×5) in the heavy fertilizer application test, ‘Darumadattan’ showed very strong lodging resistance. Heavy fertilization increase neither plant height nor seed yield, but resulted in an increase in the 1000 seed weight and a decrease in the number of nodes and rutin content in both cultivars.
Table 5.

Result of seeding density and fertilizer application tests

Fertilizer application levelaTreatmentDays to maturity daysPlant height cmNumber of nodes on main stem No./plantNumber of branches on the main stem No./plantDegree of lodgingbTotal dry weight kg/aSeed yield kg/a1000 seed weight gRutin contents mg/100 g DW
Interrow space cmSeeding density seeds/m2DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8DarumadattanHokkai T8
Standard305284766313522.619.47.95.70440.037.924.216.814.819.415921984
Standard3010484756012722.418.26.83.60334.040.220.018.414.617.717512079
Standard3020881735410720.517.04.62.90327.029.115.514.414.021.017141998
Standard602684786013822.220.77.37.00329.137.217.617.715.219.018252176
Standard605282746312320.418.46.45.40330.525.316.012.414.918.616442125
Standard6010481745411419.417.54.73.90322.832.113.216.614.420.116312074
ANOVACultivar (C)******NSNS****
Treatment (T)*****NSNSNSNS
C × TNSNSNSNSNSNSNS
Standard305284766313522.619.47.95.70440.037.924.216.814.819.415921984
Standard602684786013822.220.77.37.00329.137.217.617.715.219.018252176
Heavy305289825413320.119.98.26.90539.943.122.920.315.520.716872093
Heavy602688845712622.418.58.57.70430.930.218.515.816.221.317122185
ANOVACultivar (C)*****NSNS****
Treatment (T)NSNSNS*NSNS*
Fertilizer (F)NS*NSNSNS**NS
C × TNSNSNSNSNSNSNS
C × FNSNSNSNSNS*NS
T × FNSNSNSNSNSNSNS
C × T × FNS*NSNSNSNSNS

Standard: N-P-K = 0.28-0.28-0.28 kg/a, Heavy: 5 times of standard, N-P-K = 1.40-1.40-1.40 kg/a.

0 none–5 completely lodged.

*, **, NS: significantly different at the 0.05 and 0.01 levels and non-significant at the 0.05 level.

Discussion

In this study, we found that ‘Darumadattan’ plants were shorter than ‘Hokkai T8’ plants (Tables 3, 5). The main stem length (the sum of the internode lengths) was also shorter in ‘Darumadattan’ (data not shown); this confirms the results of Kasajima , who measured the main stem length in ‘Darumadattan’ (IRBFT-20, in the test) plants when the line was in the process of registration. The numbers of nodes on the main stems of the two cultivars were similar (Tables 3, 4). The internode elongation pattern in ‘Darumadattan’ (IRBFT-20) was examined in detail by Kasajima . Since each internode was shorter in ‘Darumadattan’, the main stem was also shorter than in the original cultivar. On the other hand, although ‘Darumadattan’ revealed latter maturity and lower 1000 seed weight than those of ‘Hokkai T8’, the effect of the semidwarf gene for these characteristics is unknown. As the semidwarf trait in ‘Darumadattan’ (‘IRBFT-20’) is controlled by a single nuclear recessive gene sdA (Morishita ), this trait could be easily introduced into other cultivars by crossing using the hot-water emasculation method (Mukasa ). A further semidwarf cultivar is currently under development (Suzuki 2019). In the seeding density and heavy fertilizer application tests, the seed yield of ‘Darumadattan’ was higher than that of ‘Hokkai T8’ except when the seeding density was 104 seeds/m2 and the interrow space was 60 cm (Table 5). A fertilizer application test conducted in Hokkaido by Kasajima resulted in no considerable differences in seed yield between the two cultivar genotypes. Depending on the cultivation year and site, the seed yield of ‘Darumadattan’ was sometimes lower and sometimes higher than that of ‘Hokkai T8’ (Tables 3, 5). Morishita reported a significant correlation between yield and plant height in Tartary buckwheat cultivars. Contrary to our expectations based on their results, ‘Darumadattan’ did not show a decreased yield (except when it was tested in Hokkaido) despite its reduced height. Furthermore, ‘Darumadattan’ showed sufficient yield in each test location for June and August seeding, and thus the semidwarf trait of ‘Darumadattan’ seemed to be available breeding material for various locations. The main stem diameter was 8.5 mm in ‘Darumadattan’ and 6.9 mm in ‘Hokkai T8’ (Table 3). Hagiwara showed that the correlation coefficient (negative) between the main stem diameter at the basal internode and the lodging degree was the second highest after the correlation between the main stem length and lodging resistance and was significant in 89 strains of Tartary buckwheat collected from around the world. In our study, the main stem of ‘Darumadattan’ was significantly thicker than that of ‘Hokkai T8’, the original cultivar. The increased main stem diameter is one of the reasons for ‘Darumadattan’ being a highly lodging-resistant cultivar. As mentioned above, the lodging resistance of ‘Darumadattan’ is extremely strong (Tables 3, 5, Fig. 2). Before the typhoon No. 18 in 2014, both ‘Darumadattan’ and ‘Hokkai T8’ were standing upright (Fig. 2A). However, after the typhoon No. 18, ‘Darumadattan’ plants were still standing upright (Fig. 2B), while ‘Hokkai T8’ plants were not (Fig. 2B). From photograph (Fig. 2), the lower leaves of ‘Darumadattan’ are large and expansive widely, while the upper leaves are small. It is thought that the plant shape of a low center of gravity position contributed to lodging resistance. The result of the performance test in Nagano provides further evidence of the strong lodging resistance of ‘Darumadattan’, since no lodging occurred despite strong winds, typhoon No. 18 and 19 being experienced in Nagano (Fig. 1). On the other hand, in Hokkaido in 2009, as there was no typhoon damage during the cultivation period, the season appeared to be favorable for ‘Hokkai T8’.
Fig. 2.

Photograph of ‘Darumadattan’ and ‘Hokkai T8’ before and after typhoon. Comparison of the new cultivar ‘Darumadattan’ and the original cultivar ‘Hokkai T8’. Photograph (A) was taken on September 30, 2014 (before typhoon No. 18, maximum instantaneous wind velocity was 16.5 m/s on October 6). Photograph (B) was taken on October 7, 2014 (after typhoon No. 18). ‘Hokkai T8’ lodged (the supporting cord was stretched; (B), right), whereas ‘Darumadattan’ did not (B). Photographs were taken in the field at the IRB in Ibaraki. Fertilizer applied with N, P2O5 and K2O at 0.28, 0.28, and 0.28 kg/a, respectively.

In this report, we have described the development of a semidwarf cultivar ‘Darumadattan’ from 100 seeds of ‘Hokkai T8’ irradiated by 500 Gy of gamma rays. In addition, other gamma ray irradiation (0–500 Gy) to two genetic resources and three breeding lines were also performed (Morishita ). Totally, about 4,000 seeds were provided and two semidwarf materials, one being ‘IRBFT-6’ from ‘Pontivy’ irradiated by 50 Gy and the other ‘IRBFT-20’ (Darumadattan) from ‘Hokkei 1’ (Hokkai T8) (Morishita ). Consequently, a few thousand seeds are necessary for inducing desired material by mutation breeding. As lodging reduces the Tartary buckwheat harvest, lodging resistance is a desired agronomic trait in Tartary buckwheat cultivation (Hagiwara , Inoue ). The cultivar reported here is the first promising cultivar with lodging resistance.

Author Contribution Statement

AS, HY, KD and TM designed this study and bred ‘Darumadattan’. TM conducted the gamma ray irradiation and select the semidwarf mutant. AS and TM carried out performance tests and component analysis. AS and TM drafted the manuscript. All authors read and approved the final manuscript.
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