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Set of DTH2 genes for controlling heading stage of paddy rice and haplotypes and application thereof

A technology of haplotype and heading stage, applied in the field of genetic engineering

Inactive Publication Date: 2012-07-18
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no gene cloned by QTL has been found to belong to the type that promotes flowering under long-day conditions (Tsuji H, Taoka KI and Shimamoto K (2010) Current Opinion in Plant Biology 14:1-8)

Method used

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  • Set of DTH2 genes for controlling heading stage of paddy rice and haplotypes and application thereof
  • Set of DTH2 genes for controlling heading stage of paddy rice and haplotypes and application thereof
  • Set of DTH2 genes for controlling heading stage of paddy rice and haplotypes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Construction of DTH2 near-isogenic line

[0029] 1. QTL detection, backcross and selection of DTH2

[0030] Using the RILs population of Asominori and IR24, our previous study detected multiple loci controlling heading date, one of which, DTH2, contained DTH2-a under natural long-day (NLD) conditions plants heading earlier than plants containing DTH2-i. Using the CSSLs population with Asominori as the background parent and IR24 as the donor parent, we found that CSSL18 has a significantly delayed maturation and stable expression compared with the background parent Asominori, because CSSL18 only inserts IR24 in some fragments compared with the background parent Asominori Therefore, the inserted indica fragment IR24 is the root cause of the extremely late maturity of CSSL18. We crossed CSSL18 with Asominori once, backcrossed 4 times consecutively, and then selfed to construct a CSSL18 / Asominori BC containing 374 individuals 4 f 2 population, from which DTH2 ...

Embodiment 2

[0036] Example 2 Fine mapping and map-based cloning of DTH2

[0037] 1. Identification of heading date phenotypes of two parents

[0038] Heading date refers to the date when the first ear of a single plant is pulled out; days of heading date refers to the number of days from sowing to the first ear of a single plant; extreme late heading single plant refers to the same or later heading date than the parent NIL (DTH2) Single plant; extremely late heading family refers to a family in which most of the individual plants heading in a random family are the same or later than the parent NIL (DTH2). Under NLD conditions in Beijing (Dongmen Net Room, Institute of Crop Science, Chinese Academy of Agricultural Sciences), NIL(DTH2) heading 7.4 days later than Asominori (Table 1, figure 2 C). However, under natural short-day (NSD) conditions in Hainan (Southern Propagation Base of Nanjing Agricultural University in Lingshui, Hainan), there was no significant difference in earing betwe...

Embodiment 3

[0047] Example 3 Transgenic functional complementation experiment of DTH2

[0048] 1. Construction of genome full-length complementary vector

[0049] According to the predicted candidate gene sequence, using the Nipponbare (English name Nipponbare, a rice variety with complete genome sequencing) sequence as a template, primers were designed to sequence the DTH2 genome from Asominori, and it was found that their sequences were consistent in this segment . Then, using the Nipponbare BAC clone OsJNBa33B21 (purchased from the National Gene Research Center of the Chinese Academy of Sciences) as a template, the whole genome of DTH2 was amplified (see Table 4 for primers) to obtain a 7867bp DNA fragment. This fragment was recombined into pCAMBIA1305.1 (a publicly reported and used plasmid from Australia, image 3 ) Sma I (purchased from Fermentas, Canada) site.

[0050] 2. Agrobacterium-mediated genetic transformation of rice

[0051] The obtained recombinant plasmid with correc...

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Abstract

The invention belongs to the technical field of genetic engineering, and relates to a set of DTH2 genes for controlling the heading stage of paddy rice and haplotypes and application thereof. The gene is coded with a CCT / B-box zinc finger protein, and belongs to the CONSTANS-Like transcription factor gene family, the dominant allele sequence is shown as SEQ ID NO.1, and flowering is promoted only under the long sunshine condition. The DNA sequence of the recessive allele is shown as SEQ ID NO.3. A transgenosis experiment proves that the gene has the function of advancing the heading stage of paddy rice. By sequencing 79 pieces of cultivated rice and 48 pieces of wild rice, five-base difference is discovered in the DTH2 coding region, as a result, three amino acids change, and five types of haplotypes are generated. Moreover, the transgenosis experiment proves that the gene is selected in domestication and originated in China.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a group of DTH2 genes for controlling the heading stage of rice, haplotypes and applications thereof. technical background [0002] Heading date is an important agronomic trait and may have been selected during crop domestication (Izawa T. Journal of Experimental Botany 58:3091-3097 (2007)). Cultivated rice is currently distributed over a wide area from 53° north latitude to 40° south latitude worldwide (Chang T.T. Euphytica 25, 425-441 (1976)). However, Oryza commonis, the ancestor of cultivated rice in Asia, is mainly distributed in South Asia, Southeast Asia and northern Australia, and the northernmost limit of its distribution is Dongxiang wild rice at 28°N latitude. An important purpose of rice domestication is to produce rice varieties suitable for cultivation in various regions, thereby expanding the cultivated area. The main limiting factor affec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415A01H5/00
Inventor 万建民吴玮勋陆广文江玲郑晓明钟铮铮
Owner NANJING AGRICULTURAL UNIVERSITY
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