Rapid intracellular acidification is a plant defense response countered by the brown planthopper

The brown planthopper (BPH) is the most destructive insect pest in rice. Through a stylet, BPH secretes a plethora of salivary proteins into rice phloem cells as a crucial step of infestation. However, how various salivary proteins function in rice cells to promote insect infestation is poorly understood. Among them, one of the salivary proteins is predicted to be a carbonic anhydrase (Nilaparvata lugens carbonic anhydrase [NlCA]). The survival rate of the NlCA-RNA interference (RNAi) BPH insects was extremely low on rice, indicating a vital role of this salivary protein in BPH infestation. We generated NlCA transgenic rice plants and found that NlCA expressed in rice plants could restore the ability of NlCA-RNAi BPH to survive on rice. Next, we produced rice plants expressing the ratiometric pH sensor pHusion and found that NlCA-RNAi BPH induced rapid intracellular acidification of rice cells during feeding. Further analysis revealed that both NlCA-RNAi BPH feeding and artificial lowering of intracellular pH activated plant defense responses and that NlCA-mediated intracellular pH stabilization is linked to diminished defense responses, including reduced callose deposition at the phloem sieve plates and suppressed defense gene expression. Given the importance of pH homeostasis across the kingdoms of life, discovery of NlCA-mediated intracellular pH modulation uncovered a new dimension in the interaction between plants and piercing/sucking insect pests. The crucial role of NlCA for BPH infestation of rice suggests that NlCA is a promising target for chemical or trans-kingdom RNAi-based inactivation for BPH control strategies in plants.