SP1调控碳水化合物分配对穗形态的影响

doi:10.13560/j.cnki.biotech.bull.1985.2022-1301

摘要/Abstract

摘要：

探讨SP1在水稻“源-库”系统中的调控作用，为解析稻穗形成的机理和青贮饲料稻分子育种提供科学参考。以sp1与WT为材料，进行氮处理和生长分析，表型鉴定、碳水化合物测定和RT-qPCR检测。结果发现，氮处理10 d时，sp1与WT无表型差异，但在高、中氮条件下，sp1相对生长速率显著提高；24 d时sp1叶鞘、根、植株干重提高，叶鞘氮含量降低、碳/氮比值提高，说明充裕的氮能促进sp1生长，SP1可能影响叶鞘碳-氮平衡。抽穗期sp1株高显著降低，分蘖数显著提高，稻穗变小并伴随高位分蘖发生。孕穗期sp1茎、鞘中碳水化合物含量与WT差异较小，但抽穗期茎中淀粉、蔗糖和叶鞘中淀粉、蔗糖、葡萄糖和果糖含量明显提高。SP1在叶鞘基部高表达，推测突变SP1能抑制蔗糖由茎鞘向穗的运输，导致高位分蘖发生和稻穗变小。SP1可应用于青贮饲料稻分子育种。

关键词: 穗形态, SP1, 碳水化合物, 叶鞘, 青贮饲料稻

Abstract:

In order to explore the regulation of SP1 in the rice “source-sink” system and provide scientific reference for understanding the mechanism of rice panicle formation and molecular breeding of whole-rice silage, sp1 and WT were selected as materials for nitrogen treatment and plant growth analysis, phenotype identification, carbohydrate determination, and RT-qPCR detection. The results indicated that there was no phenotypic difference between sp1 and WT at 10 d after nitrogen treatment. However, sp1 showed significantly higher plant relative growth rate than WT under high and medium nitrogen treatment. Meanwhile, sp1 demonstrated higher value of the dry weight of leaf sheath, root and plant than WT at 24 d after nitrogen treatment, while a decrease in the content of nitrogen and an increase in the value of C/N ratio in leaf sheath. It was speculated that the abundant nitrogen improved plant growth of sp1, but the nitrogen stress impaired carbon-nitrogen balance in the leaf sheath. At heading stage, sp1 showed a significant decrease in plant height and a significant increase in tiller number, and shorter panicle with the occurrence of high-node tillering. At booting stage, there was little difference of the carbohydrates content in the stem and sheath between sp1 and WT. But, sp1 showed higher content of starch, sucrose in stem, and higher content of starch, sucrose, glucose and fructose in the leaf sheath at heading stage. SP1 highly expressed at the base of leaf sheath, which implied that the mutation of SP1 would inhibit the sucrose transport from the stem and sheath to the panicle and induce high-node tillering and short panicle. Rice SP1 gene is a candidate gene for molecular breeding of whole-rice silage.

Key words: panicle architecture, SP1 gene, carbohydrate, leaf sheath, whole-rice silage

梁成刚, 汪燕, 李天, 大杉立, 青木直大. SP1调控碳水化合物分配对穗形态的影响[J]. 生物技术通报, 2023, 39(5): 152-159.

LIANG Cheng-gang, WANG Yan, LI Tian, OHSUGI Ryu, AOKI Naohiro. Effect of SP1 on Panicle Architecture by Regulating Carbohydrate Remobilization[J]. Biotechnology Bulletin, 2023, 39(5): 152-159.

图/表 7

表1 RT-qPCR分析使用的基因引物

Table 1 The primers of genes for RT-qPCR analysis

基因Gene	正向引物 Forward primer（5'-3'）	反向引物 Reverse primer（5'-3'）
SP1	GTTCGAACCGCACGTCTAGT	GGGGACTCATATACATCCACCC
UBI	GGAGCTGCTGCTGTTCTTGG	CACAATGAAACGGGACACGA

图1 氮处理10 d时植株表型（A）与氮处理10-20 d相对生长速率（B） HN：高氮处理；MN：中氮处理；LN：低氮处理。*与**分别表示差异达到显著（P<0.05）或极显著（P<0.01）水平，下同。标尺为10 cm

Fig. 1 Plant phenotype at 10 d after nitrogen treatment （A）, and the relative growth rate from 10 d to 20 d after nitrogen treatment（B） HN: High nitrogen treatment; MN: middle nitrogen treatment; LN: low nitrogen treatment. * and ** indicate significant（P<0.05）or extremely significant（P<0.01）difference. The same below. Scale bar=10 cm

表2 不同氮浓度下的水稻农艺性状

Table 2 Agronomic traits of rice under different nitrogen conditions

材料 Material	氮处理 Nitrogen treatment	根重 Root weight/mg	叶鞘重 Leaf sheath weight/mg	叶重 Leaf weight/mg	植株干重 Plant dry weight/mg
WT	HN-10 d	33.94±0.54	38.66±1.44	43.56±0.76	116.16±2.12
sp1	HN-10 d	35.66±1.17	43.96±0.83*	44.90±0.82	124.52±1.08**
WT	MN-10 d	38.18±1.37	41.78±1.05	40.98±1.56	120.94±2.54
sp1	MN-10 d	38.14±0.78	45.52±0.91*	40.32±1.55	123.98±2.05
WT	LN-10 d	37.76±1.09	37.64±1.16	32.06±1.33	107.46±1.23
sp1	LN-10 d	42.84±1.22	39.74±0.61	34.58±0.54*	117.16±1.33**
WT	HN-24 d	106.84±3.70	210.38±8.05	218.10±14.36	535.32±12.39
sp1	HN-24 d	145.24±11.94*	294.80±27.57*	241.12±19.97	681.16±31.42**
WT	MN-24 d	152.26±11.37	213.24±13.75	164.36±3.92	529.86±13.56
sp1	MN-24 d	194.00±19.20*	297.76±33.23*	212.24±18.34**	737.86±37.61**
WT	LN-24 d	126.44±5.50	170.76±5.10	104.60±1.64	401.80±7.08
sp1	LN-24 d	144.42±2.64*	193.42±4.76*	99.26±7.14	437.10±5.15**

表3 不同氮浓度处理24 d植株各器官中的碳氮百分含量

Table 3 Carbon and nitrogen content in rice plant organs at 24 d under nitrogen treatment

处理/材料 Treatment/ Material	氮含量Nitrogen content/%			碳含量Carbon content/%			碳/氮比Carbon/Nitrogen ratio
处理/材料 Treatment/ Material	根 Root	叶鞘 Leaf sheath	叶片 Leaf blade	根 Root	叶鞘 Leaf sheath	叶片 Leaf blade	根 Root	叶鞘 Leaf sheath	叶片 Leaf blade
HN-WT	1.68±0.05	2.07±0.01	3.79±0.04	45.26±1.00	40.99±0.20	43.54±0.39	27.10±1.24	19.77±0.20	11.48±0.10
HN-sp1	1.68±0.02	1.90±0.04*	3.71±0.03	43.80±0.12	40.82±0.10	43.15±0.03	26.09±0.35	21.54±0.42*	11.63±0.10
MN-WT	1.12±0.04	1.32±0.04	3.17±0.06	45.00±0.29	41.33±0.06	43.38±0.07	40.41±1.37	31.48±0.20	13.70±0.10
MN-sp1	1.07±0.02	1.28±0.02	2.97±0.07	43.59±0.35	40.86±0.10*	42.89±0.12	40.65±0.87	31.87±0.42	14.48±0.10
LN-WT	0.86±0.03	1.08±0.00	2.67±0.06	47.31±0.38	43.09±0.32	42.95±0.16	55.63±2.47	39.94±0.44	16.14±0.38
LN-sp1	0.79±0.04	0.89±0.01**	2.46±0.06	48.32±0.46	44.20±0.21	43.51±0.33	61.76±2.88	49.52±0.63**	17.75±0.36

图2 水稻sp1的表型特征与SP1的RT-qRCR分析 A：水稻野生型与sp1植株的表型。红色箭头指向sp1高位分蘖，蓝色箭头指向sp1稻穗；B：水稻野生型与sp1的谷粒性状；C：SP1的表达量分析。LB：叶片，TLS：叶鞘上部，BLS：叶鞘基部

Fig. 2 Phenotype of rice sp1 and RT-qRCR analysis of SP1 gene A: Plant of WT and sp1. Red arrows point to high-node tillerings of sp1. Blue arrow points to panicle of sp1. B: Cereals of WT and sp1. C: RT-qRCR analysis of SP1 gene. LB: Leaf blade. TLS: Top of leaf sheath. BLS: Base of leaf sheath

图3 移栽后sp1顶端全展叶叶绿素相对含量SPAD值（A）、株高（B）与有效分蘖（C）的动态变化

Fig. 3 Dynamic changes of SPAD value in the top comple-tely-expanded leaf（A）, plant height（B）and effective tiller number（C）of sp1 after transplanting

图4 移栽后70 和80 d水稻sp1顶端组织的淀粉（A）、蔗糖（B）、葡萄糖（C）和果糖（D）含量 C2：穗下第2-3节间茎；C3：穗下第3-4节间茎；LS2：倒2叶鞘；LS3：倒3叶鞘

Fig. 4 Contents of starch（A）, sucrose（B）, glucose,（C）and fructose（D）in the upper tissues of sp1 collected at 70 d and 80 d after transplanting C2: The internode culm between 2nd and 3rd below panicle. C3: The internode culm between 3rd and 4th below panicle. LS2: The second top leaf sheath. LS3: The third top leaf sheath

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