Development of Heat Tolerant Two-Line Hybrid Rice Restorer Line Carrying Dominant Locus of OsHTAS

doi:10.1016/j.rsci.2020.11.011

Abstract

Abstract:

In order to create novel germplasm resources for breeding heat tolerant variety, we transferred a dominant allele OsHTAS, previously characterized and cloned from a high-temperature tolerant local variety HT54, which was collected from the rice production area of southern China, into a high- temperature sensitive intermediate breeding line HT13 through six rounds of successive backcross by using marker-assisted selection. The molecular analysis showed that the recovery of genetic background of a resultant near isogenic line (NIL), MHT13, was around 99.8%. The OsHTAS gene introduced in the MHT13 expressed normally in the HT13 genetic background, mediating heat tolerance and phenotype similar to those of the donor parent HT54. The major agronomic traits of MHT13 resembled those of the recurrent parent HT13. Moreover, MHT13 had high general combining ability and its rice quality reached the grade 3 standard of edible high-quality rice issued by Ministry of Agriculture of the People’s Republic of China, which greatly improved its application value in rice production.

Key words: rice, heat tolerance, high temperature, yield, near isogenic line, marker-assisted selection, grain quality, OsHAT gene

Jan Mehmood, Shah Gulmeena, Yuqing Huang, Xuejiao Liu, Peng Zheng, Hao Du, Hao Chen, Jumin Tu. Development of Heat Tolerant Two-Line Hybrid Rice Restorer Line Carrying Dominant Locus of OsHTAS[J]. Rice Science, 2021, 28(1): 99-108.

Figures/Tables 6

Fig. 1. Schematic diagram of marker-assised backcross breeding and graphical genotype of near isognic line MHT13.A, Flow chart of marker-assisted backcross breeding for developing near isogenic line with the dominant locus of OsHTAS. FS, Foreground selection; BS, Background selection.B, Graphical genotype of MHT13. The purple solid bar represents the foreground fragment containing OsHTAS introduced from HT54, while the hollow vertical bar represents the background genotype inherited from HT13. The background selection was carried out using 144 SSR markers. Chr, Chromosome.

Table 1 Agronomic performance of MHT13 under normal growth conditions (Hangzhou, 2017).

Line	Growth duration (d)	Plant height (cm)	No. of tillers per plant	Panicle length (cm)	No. of grains per panicle	1000-grain weight (g)	Spikelet fertility (%)
HT54	96.0 ± 0.0	85.2 ± 1.7	8.0 ± 1.4	20.8 ± 1.4	119.6 ± 5.4	28.4 ± 0.9	89.8 ± 1.7
HT13	126.0 ± 0.0	93.1 ± 2.5	7.8 ± 0.9	23.5 ± 0.8	115.1 ± 7.0	24.2 ± 0.6	93.4 ± 2.6
MHT13	126.0 ± 0.0	92.8 ± 1.0	8.0 ± 1.2	23.8 ± 0.9	116.8 ± 9.7	24.3 ± 1.0	92.2 ± 2.9

Table 2 Agronomic performance of MHT13-derivative hybrids under field conditions in 2017 and 2018, respectively.

Cross	Growth duration (d)	Plant height (cm)	No. of tillers per plant	Panicle length (cm)	No. of grains per panicle	1000-grain weight (g)	Seed-setting rate (%)	Yield per plant (g)
2017, Hangzhou, China
218S/HT13	134.2 ± 1.2 a	127.5 ± 5.2 a	14.6 ± 0.1 a	26.2 ± 1.5 a	236.8 ± 10.5 a	27.5 ± 0.2 a	84.8 ± 4.0 a	58.83 ± 0.10 b
218S/MHT13	134.0 ± 1.1 a	127.8 ± 2.6 a	15.5 ± 1.5 a	27.6 ± 2.1 a	235.3 ± 13.0 a	27.0 ± 1.0 a	87.4 ± 1.4 b	65.90 ± 1.20 a
208S/HT13	135.7 ± 1.3 a	124.8 ± 0.9 a	15.2 ± 0.5 a	25.9 ± 2.5 a	218.8 ± 5.4 a	24.5 ± 3.1 a	85.2 ± 2.0 a	66.20 ± 0.57 b
208S/MHT13	135.7 ± 1.2 a	125.8 ± 1.9 a	16.0 ± 1.0 a	26.1 ± 0.5 a	227.4 ± 3.5 b	26.5 ± 0.5 a	86.8 ± 5.0 a	70.36 ± 0.28 a
2018, Changxing, China
218S/HT13	134.3 ± 2.3 a	127.0 ± 2.2 a	13.8 ± 1.5 a	27.1 ± 1.1 a	243.1 ± 9.3 a	26.2 ± 1.5 b	88.0 ± 5.6 a	64.77 ± 1.30 c
218S/MHT13	134.2 ± 1.8 a	127.0 ± 0.5 a	15.5 ± 0.7 b	26.4 ± 2.2 a	244.4 ± 5.0 a	27.4 ± 2.0 a	89.5 ± 2.2 a	72.03 ± 0.60 a
228S/HT13	133.2 ± 2.1 a	125.4 ± 1.5 a	15.3 ± 0.7 a	25.5 ± 3.4 a	221.2 ± 12.1 a	27.9 ± 2.2 a	73.3 ± 0.6 a	61.52 ± 0.85 a
228S/MHT13	133.4 ± 1.6 a	130.2 ± 3.4 b	14.7 ± 0.3 a	26.1 ± 2.0 a	220.4 ± 2.1 a	28.4 ± 0.3 a	75.7 ± 3.3 a	63.77 ± 0.43 a

Fig. 2. Dynamic expression, phenotype and heat tolerance mechanism of OsHTAS in MHT13 and its derivative hybrids.A, Dynamic expression features of OsHTAS under 48 ºC high temperature.B, Expression changes of OsHTAS in hybrids of 208S/HT13 and 208S/MHT13 before and after 48 ºC high temperature treatment.C, Changes of seed-setting rate of heat resistant and sensitive parents and their continuously-backcrossing obtained MHT13 and negative materials before and after 38 ºC high temperature treatment.D, Changes of seed-setting rate of heat-sensitive HT13 and its improved MHT13-formulated hybrids before and after 38 ºC high temperature treatment.E, ABA content of MHT13 and its parental lines before and after 48 ºC high temperature treatment.F, Changes of water loss before and after 48 ºC high temperature treatment.High temperature (48 ºC) treatment lasting 79 h was carried out at the seedling stage, and heat stress response was then scored and photographed after end of treatment and recovery for 10 d. High temperature (38 ºC) treatment was carried out on the day of flowering, the treatment lasted 6 h and the seed-setting rate was counted after maturity. Error bars indicate SE based on three biological replicates. Significant differences between lines were determined by the Duncan’s multiple range test at the 0.05 level.

Fig. 3. High temperature tolerance of MHT13 at seedling stage. Neg represents the negative line obtained by marker-assisted breeding. The treatment used was heat stress at 48 ºC for 79 h.

Table 3 Evaluation results of grain quality of MHT13 according to quality standard of edible indica rice issued by the Ministry of Agriculture of China.

Index	BRR (%)	HMRR (%)	GL (mm)	CD (%)	TD (Rank)	ASV (Rank)	GC (mm)	AC (%)	MRR (%)	GLWR	CGR (%)	PC (%)
Grade 3 standard	≥ 77.0	≥ 52.0	> 6.5, 5.6‒6.5 or < 5.6 ^a	≤ 5.0	≤ 2	≥ 5.0	≥ 50	13.0-22.0	-	-	-	-
Test result	80.6	64.3	6.7	3.3	2	7.0	58	15.0	72.3	3.3	13	8.36
Single judgment	2	1	Long grain	3	2	1	3	1	-	-	-	-

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