Scientia Agricultura Sinica ›› 2019, Vol. 52 ›› Issue (7): 1227-1236.doi: 10.3864/j.issn.0578-1752.2019.07.010

• PLANT PROTECTION • Previous Articles     Next Articles

Effects of Oxytetracycline Treatment on the Control of Sweet Orange Huanglongbing and the Expression of PP2 Gene

YAO TingShan1,2,ZHOU Yan1,DIANN ACHOR3,WANG Nian3,ZHOU ChangYong1,2()   

  1. 1 Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing 400712, China
    2 College of Plant Protection, Southwest University, Chongqing 400716, China
    3 Citrus Research and Education Center, University of Florida, Lake Alfred, Florida 33850, USA
  • Received:2018-11-23 Accepted:2019-01-23 Online:2019-04-01 Published:2019-04-04
  • Contact: ChangYong ZHOU E-mail:zhoucy@cric.cn

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Abstract:

【Objective】The objective of this study is to evaluate the control efficacy of oxytetracycline (OTC) on sweet orange Huanglongbing (HLB), detect the expression of phloem protein 2 (PP2) gene, and to provide a scientific basis for the effective prevention and control of HLB as well as a reference for OTC mechanism research. 【Method】OTC, which can effectively inhibit and kill pathogenic bacteria, was injected with 0.1 g/tree of 4-year-old Valencia sweet orange with different severity of HLB (newly infected, mild infected and severely infected plants), and then the contents of Cadidatus Liberibacter asiaticus (Las), starch and PP2 gene of leaves were evaluated at 7, 30, 60, 90 days post injection, respectively. Furthermore, 90 days after injection, the young and mature tissues of the plants were observed by using starch staining light microscopy (LM). The extraction of tender leaves was investigated 80 days after OTC injection. 【Result】Trunk injection of OTC 0.1 g/tree had the most obvious therapeutic effect on newly infected Valencia sweet orange. Seven days after injection, Las could not be detected and it maintained for 90 days. The starch content in mature leaves decreased obviously, but starch grains were still enriched in stems. The Las content of mild infected Valencia leaves decreased from (1.68×10 6±858884) cells/g leaf to (7.21×10 4±30981) cells/g leaf, the descending range was obvious, and the starch content in mature leaves showed a decreasing trend within 90 days. The Las content of severely infected Valencia leaves decreased from (4.10×10 8±3.04×10 8) cells/g leaf to (2.80×10 7±2.70×10 7) cells/g leaf, but the starch content in mature tissues of severely infected plants was still high, except the newly autumn shoots. The results indicated that 0.1 g/tree OTC was not enough to cure 4-year-old severely infected Valencia. The expression of PP2 gene in Valencia significantly decreased 30 days after OTC injection, and remained stable in the subsequent 90 days, which was consistent with the expression level at 30 days after injection.【Conclusion】OTC can be used in the prevention and control of HLB, and has better therapeutic effect on newly infected Valencia (Las content is less than 9.00×10 5 cells/g leaf) and mild infected Valencia (Las content is 9.00×10 5-9.00×10 7 cells/g leaf). It was not suitable for the treatment of severely infected plants (Las content is more than 9.00×10 7 cells/g leaf). After injection of OTC, the expression of PP2 gene decreased obviously, suggesting that OTC can effectively reduce the stress of phloem pathogens.

Key words: Huanglongbing (HLB), Cadidatus Liberibacter asiaticus (Las), oxytetracycline (OTC), starch, PP2

Table 1

Grouping based on Las content in leaves of Valencia plants"

编号
Number		 植株
Plant		 Las含量
Las content
(cells/g leaf)		 编号
Number		 植株
Plant		 Las含量
Las content
(cells/g leaf)		 编号
Number		 植株
Plant		 Las含量
Las content (cells/g leaf)
1 初感染-T
Newly infected-T		 1.58×105 7 轻度发病-T
Mild infected-T		 2.60×106 13 重度发病-T
Severely infected-T		 4.60×108
2 1.32×105 8 1.54×106 14 3.59×108
3 1.23×105 9 9.04×105 15 4.10×108
平均
Average		 1.38×105 平均
Average		 1.68×106 平均
Average		 4.10×108
4 初感染-CK
Newly infected-CK		 1.01×105 10 轻度发病-CK
Mild infected-CK		 2.10×107 16 重度发病-CK
Severely infected-CK		 4.31×108
5 1.21×105 11 4.59×106 17 2.60×108
6 9.34×104 12 9.05×106 18 7.19×108
平均
Average		 1.07×105 平均
Average		 1.16×106 平均
Average		 4.70×108

Fig. 1

The change of Las content in leaves of Valencia with HLB after OTC trunk injection Different lowercases indicate significant difference (P<0.05). The same as below"

Fig. 2

The change of starch content in leaves of Valencia with HLB after OTC trunk injection"

Fig. 3

Light micrographs of Valencia tissue cross sections before and after OTC treatment A-F: Contrast charts of newly infected Valencia before and after OTC injection. Among them, A is the mature leaf before treatment; B is the mature leaf after treatment 90 days; C is the tender leaf before treatment; D is the tender leaf after treatment 90 days; E is the stem before treatment; F is the stem after treatment 90 days. G-L: Contrast charts of severely infected Valencia before and after OTC injection. Among them, G is the mature leaf before treatment; H is the mature leaf after treatment 90 days; I is the tender leaf before treatment; J is the tender leaf after treatment 90 days; K is the stem before treatment; L is the stem after treatment 90 days. M-O: Healthy Valencia plant control. Among them, M is healthy mature leaf; N is healthy young leaf; O is healthy stem. Non-lignified/cellulose cell walls are stained red or purple, cytoplasm blue, and starch granules reddish yellow. The blue spots indicated by blue arrows represent phloem plugging. The reddish yellow granules indicated by yellow arrows represent starch. Co: Cortex; Fi: Fiber; P: Phloem; Pi: Pith; X: Xylem"

Fig. 4

The change of new shoot (leaf) of Valencia after OTC trunk injection"

Fig. 5

The change of PP2 gene expression of Valencia with HLB after OTC trunk injection"

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