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Performance of Some Pi40 Gene Derived Japonica

Breeding Lines for Blast Disease Resistance in Turkey

 Sürek, H1*., Beşer, N2., Del Valle M. M3., Jena, K.K3.

1Trakya Agricultural Research Institute, Edirne, Turkey

2Trakya University, Engineering Faculty, Department of Genetic and

Bioengineering, Edirne, Turkey.

3Novel Gene Resources Laboratory, Plant Breeding Division, International Rice Research Institute, DAPO Box 7777, 1226 Metro, Manila, Philippines.

 *Corresponding author: Email: This email address is being protected from spambots. You need JavaScript enabled to view it.

 

Abstract

 Rice is cultivated in tropical as well as temperate ecosystems as indica and japonica cultivars, respectively. Rice still is a high profitable crop in Turkey. Rice is cultivated as a monoculture crop in summer and it increases the disease incidence risks mainly fungal blast disease caused by Magnaporthe oryzae. The most severe blast disease infection and yield losses were recorded in Trakya region of Turkey in 1995 where only half of rice was produced.  The yield losses occurred in 20% of the 25000 hectares areas in this region and also the head rice yield was drastically decreased. Application of fungicide increases the production cost and environmental concern. Using the resistant varieties is the most economical and environmental friendly way to control this disease. A broad-spectrum blast resistance gene, Pi40 is available at the International rice research Institute, Manila, Philippines. The objective of this study was to determine performance of  Pi40 blast resistant gene transferred into two Turkish cultivars (Osmancık-97 and Halilbey) through development of japonica breeding lines in a Turkey-IRRI cooperative Research Project.

             The two elite cultivars (Osmancık-97 and Halilbey) susceptible to blast were crossed with the Pi40 donor line, IR83260-1-1-1-5-B-3-1-2-B in a japonica background and followed by marker-assisted backcross breeding (MAS) at IRRI. After reaching to BC3F5 generation, advanced backcross progenies or lines were sent to TARI (Trakya Agricultural Research Institute) for blast screening at different hotspot locations and for agronomic trials. The Pi40-derived backcross lines in Osmancık-97 and Halilbey backgrounds showed resistance to blast disease at observation nurseries in 2014 and the yield trials in Turkey in 2015. The yield trials were carried out in a complete randomized block design with three replications in two locations. According to the results of  yield trials and disease tests, two Osmancık-97 lines (IR99598-2-10-1 and IR99586-3-7-14-B) and two Halilbey lines ( TR-7-3-2-1-2 and IR99599-1-12-B) showed better performance than recurrent parents in terms of rice yield and agronomic traits. They were nominated for registration as commercial variety in Turkey at the end of 2015 crop season.

             The results clearly demonstrated that the broad-spectrum resistance gene, Pi40 in the background of two elite cultivars of Turkey provided resistance against blast disease pathotypes in hotspot locations. Performance of selected advanced backcross breeding lines with the Pi40 gene showed high yield potential and blast disease resistance with superior grain quality compared to the recurrent japonica cultivars of Turkey. The information gained in this study will be highly applicable for enhancing genetic base of host-plant resistance.

Key Words: Blast, marker-assisted backcross, Pi40 gene, resistance, rice (Oryza sativa L.)

            Introduction

            Rice is cultivated in tropical as well a s temperate ecosystems as indica and japonica cultivars, respectively. Rice cultivated in limited area due to shortage of available irrigation water for rice crop in temperate regions of Turkey (Sürek, 1997).  However, rice still is a high profitable crop in Turkey. Therefore, the farmers are very willing to grow rice, it creates monoculture rice growing systems and it also increases the disease infection risks. The cultivated rice type is typical japonica (Beşer and Surek, 2012).

            All the rice diseases recorded in the other rice growing countries do not occur in Turkey. The three fungal rice diseases observed are: Blast, Brown leaf spot, Bakanea and foot rot caused by the fungus Magnaporthe oryzae, Helminthosporium oryzae, and  Fusarium moniliforme , respectively (Sürek, 1995).

            Rice blast is caused by the fungus Magnaporthe oryzae (anamorphi: Phyricularia grisea Sacc.). Blast severely effects lowland rice in temperate and subtropical areas of Asia, and is highly destructive to upland rice in tropical areas of Asia, Latin America, and Africa (IRRI). Wang et al., (2014) reported that the blast disease first reported in the United

states in 1876, and has been identified in 85 rice producing countries or regions worldwide.          In China, 40-50% yield losses were observed under severe rice blast infection; in some cases, 100% yield losses were found in severely infected fields (Zhanget al., 2010). Yield losses of 5-10%, 8%, and 14% were reported in India from 1960 to 1961, Korea from the mid-1970s, and China from 1980 to 1981, respectively (IRRI). The highest yield losses were recorded in the Philippines; ranging from 50% to 85% in 1963 (IRRI, 2014).

            The earlist attemts to study pathogenic fungi in Turkey were made by Bremer and Özkan (1946) and Göbelez (1953). They reported that  blast was the most important disease in Turkey. Gobelez also observed up to 25 to 75% yield losses in some rice fields in the Black Sea Region. On the other hand, Teknikel etal., (1980) reported sever infection up to 90% in the Meditterranean coastal areas of Turkey. 8,33% yield reduction was observed by Oran (1975) in the south-eastern part of the country.

            The most severe blast disease infection and yield losses recorded in Trakya region of Turkey in 1995. where the half of rice isproduced.  The yield losses was 20% in 25000 hectares areas in this region and also the head rice yield was drastically decreased (Sürek and Başer, 1997). Since then, the blast infection occurs depending on weather conditions, such as high temperatures, increases in rainy days, high relative humidity and lower night temperature etc. At the same time, some agricultural practices used by the farmers increase the disease infection risks. These practices are monoculture rice faming, excessive  nitrogen application, nitrogen application at  non recommended  times, late planting, and high plant density etc.

            In order to control blast disease, The Turkish rice farmers use fungicides. They apply the fungicide  several times depending on the disease infection starting time and intensity. This practice increases the production cost and environmental concern.

            Using the resistant varieties is the most economically and environmentally friendly way to control this disease. However, the majority of rice varieties used in the rice production are susceptible to the blast disease in Turkey. One of them is Osmancık-97 which is cultivated more then at 50% of total rice growing area and its quality characteristics are well accepted by The Turkish consumers. The other one with high yield potential is Halilbey, but it is highly susceptible to the panicle blast.

            Rice cultivars resistance to blast disease are developed by conventional breeding using the sources of resistance from land races of diverse rice varieties. However, the resistance breaks down in two to three years  (Miah et. al., 2013). Novel sources of blast resistance genes have been identified in the introgression lines derived from wild Oryza species (Jena and Kim, 20109.

            The Pi40 gene has been introgression line that inherited the resistance gene from a wild species O. australiensis and it has durable broad-spectrum of blast resistance (Jeung et. al., 2007). The Pi40 gene confers blast resistance at seedling as well as reproductive stages.  Suh et al., (2009) determined that the Pi40 gene provided durable resistance to blast. Practicing marker assisted backcross breeding method, also,  Lee et al., (2015) obtained panicle blast resistant variety using Pb1 resistant gene in Korea and Hua et al., (2015 introgressed Pi39 blast resistant gene into two elite cultivars in China.

            Marker-assisted backross breeding is a powerful tool to incorporate novel resistance genes into susceptible cultivars and to develop improved breeding lines in a short period of time ( Kush and Jena, 2009; Jena and Mackill, 2008).

            The objective of this study was to determine performance of  Pi40 blast resistant gene transferred Turkish cultivar (Osmancık-97 and Halilbey) backcross japonica lines in Turkey.

              Material and Methods

             Materials

             Two elite cultivars of Turkey, Osmanık-97 and Halilbey were used as the recurrent parents, and IR83260-1-1-1-5-B-3-1-2-B (4128) carrying the Pi40 gene was used as the donor parent.

             Method

             A Turkey-IRRI Cooperative Research Projects conducted to develop blast-resistant japonica rice using marker-assisted backcross breeding conducted at IRRI and TARI in Turkey. The aim of this project was to transfer the Pi40 blast resistant gene into two Turkish elit cultivars, Osmancık-97 and Halilbey. Both of these varieties are susceptible to blast disease. This project started in 2010 at IRRI, two elite cultivars of Turkey (Osmancık-97 and Halilbey) were screened for their blast reaction to virulent blast races. The two elit varieties showed susceptible reaction to blast. These varieties were crossed to the Pi40 donor in a japonica background (IR83260-1-1-1-5-B-3-1-2-B). After then, marker-assisted backcross breeding (MAS) practiced and backcross segregating materials at different generations were obtained. Also, through molecular analysis the materials or line determined with Pi40 gene at IRRI. After reaching advanced bakcross progenies or lines were sent to TARI for evaluating blast screening in different hotspot locations and for agronomic trials.

            The advanced bakcross lines showed good performance for agronomic traits and resistance to blast disease, tested in the yield trials. Three yield trials conducted in two locations in Edirne and İpsala town of Edirne (the main rice growing region in Turkey) in 2015. One yield trial consisted of Osmancık-97 backcross lines and the other consisted of Halilbey backcross lines in Edirne location at TARI. İpsala location consisted of both selected promising backcross lines of Osmancık-97 and Halilbey cultivars. The yield trials were carried out in a complete randomized block design with three replications. The recurrent parents were used as check in the experiment. Rice yield data were subjected to ANOVA using   MSTAT-C package programme. LSD was calculated for comparison of cultivar means.

            The disease test observation nurseries were conducted at the different hotspot locations. For these nurseries, first the seedlings were grown in seed beds, and then 25 days old seedlings were transplanted in one metter row, distance between rows was 15 cm and 7 cm between hills. Encourage disease infection, highly susceptible check variety's seedlings were transplanted around the plot and high rates of nitrogen fertilizer were applied. Disease evaluation was done according to Standard Evaluation Evaluation System for Rice (IRRI, 2014).

            The data were recorded for plant height, panicle length, days to flowering and maturity, the number of panicle per squarmetter, spikelet fertility, 1000 grain weight for rice and milled grains, rice and milled grain length, rough rice  and head rice yield.

             Results and Discussions

            During the past decade, nearly 50 blast resistance genes have been identified in the wold. However, most of them are race specific and resistant cultivars possessing these genes break down within 2 to 3 years. Durable resistance of gene to a disease is resistance that remains effective its prolonged and widespread use in an environment favourable to the disease. The most powerful test for the durability of blast resistance occurs when a cultivars is widely grown for a long period in an environment favouring the disease.

            Pi40 blast resistant gene transferred into two elit Turkish rice varieties, Osmancık-97 and Halilbey, using marker assisted backcross breeding method in International Rice Research Institute (IRRI). Developed backcross breeding lines were sent to Trakya Agricultural Research Institute for disease evaluation test at hotspot areas. At the same times, the lines showed resistance to disease under field conditions, they tested in observation nursery and yield trials to evaluate in term of agronomic and  quality traits. After getting the disease test and yield trial results, the selected two Osmancık-97 backcross lines and two Halilbey backcross lines were nominated for registration as commercial variety at the end of 2015. They are being tested in National Rice Variety Registration Trial in 2016.

            The disease evaluation results of two Osmancık-97 and two Halilbey backcross lines having Pi40 genes are seen in table-1. When a comparison is done between Pi40 gene carrying backcross lines and recurrent parents. The Pi40 genes backcross lines seem to be more resistant than  recurrent parents, Osmancık-97 and Halilbey  to leaf and panicle blast disease. Similar results reported by Suh et. al., (2009). On the other hand, Lee et. al., (2015) in Korea and Hua et. al., (2015) in China, using Pb1 and Pi39 blast resistant genes developed blast resistant varieties, respectively.

            The average yields and agronomic traits of backcross lines and recurrent parents are seen in table-2. The developed backcross lanes have higher rice yields and better agronomic traits than recurrent parents. That is why, these lines were nominated for registration as a commercial variety.

 Table 1. The disease evaluation results of two Osmancık-97 and two Halilbey backcross lines in hotspot locations.

 

Designation

 

Recurrent

Parent

Test

Year

Edirne İpsala

Town

Tekirdağ Hayrabolu Town

Balıkesir Gönen Town

Çanakkale

Biga Town

LB

NB

LB

NB

LB

NB

LB

NB

IR99598-2-8-10-1

Osmancık-97

2013

1

0

--

--

1

1

--

--

IR99599-1-12-B

Halilbey

2014

1

5

1

1

1

1

1

5

IR99586-3-7-14-B

Osmancık-97

1

1

1

1

1

1

1

1

IR83260-1-1-1-5-B-3-1-2-B (Pi40 gene donor)

 

1

1

1

1

1

1

1

1

Osmancık-97  (Recurrent Parent)

 

3

7

1

1

1

3

1

3

Halilbey (Recurrent Parent)

 

1

7

1

1

1

3

1

5

Diyarbakır Yerli (Susceptable Check)

 

9

9

9

9

9

9

9

9

Sarıçeltik (Susceptable Check)

 

9

9

9

9

9

9

9

9

IR99598-2-8-10-1

Osmancık-97

2015

0

0

--

--

0

0

0

0

IR99586-3-7-14-B

Osmancık-97

0

0

--

--

0

0

0

0

Osmancık-97  (Recurrent Parent)

 

7

7

--

--

3

3

3

3

TR-7-3-2-1-2

Halilbey

3

0

--

--

0

0

0

0

IR99599-1-12-B

Halilbey

0

0

--

--

0

0

0

0

Halilbey (Recurrent Parent)

 

0

5

--

--

0

3

0

3

Sarıçeltik (Susceptable Check)

 

9

9

--

--

9

9

9

9

Kızıltan (Susceptable Check)

 

9

9

--

--

3

9

9

9

LB: leaf blast infection, NB: node blast infection (including node, neck, and panicle blast).

 

Table 2. The main characteristics of Osmancık-97 and Halilbey backcross lines nominated for registration.

Designation

The average yield (ton/ha)

Plant height

cm

 

Panicle length

Cm

Days

 to flowering

Days

 to maturity

The number of panicle

m-2

Spikelet

fertility

  %

1000

 rice

grain weight g.

1000 millet grain weight

g.

Head

rice

yield

  %

Rice

 grain

length

mm

Milled rice

grain length

IR99586-3-7-14-B

7,6

92,3

14,7

79

122

306

94,0

33,1

23,8

66,1

8,8

6,1

IR99598-2-8-10-1

8,1

91,9

16,7

81

127

363

91,3

31,7

23,7

61,5

8,4

6,0

Osmancık-97 (check)

7,6

98,1

15,5

81

124

385

89,9

33,6

24,6

61,6

8,8

6,3

 

IR99599-1-12-B

8,0

95,3

16,8

77

121

300

92,2

30,9

22,0

59,0

8,7

6,1

TR-7-3-2-1-2

7,7

89,2

17,8

83

132

340

76,6

32,4

23,7

64,1

8,9

6,4

Halilbey (check)

7,3

97,6

16,7

78

124

340

93,0

34,9

24,4

56,1

9,2

6,4

 


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