Cereals high in amylose content (AC) and resistant starch (RS) offer potential health benefits. Previous studies using chemical mutagenesis or RNA interference have demonstrated that starch branching enzyme (SBE) plays a major role in determining the fine structure and physical properties of starch. However, it remains a challenge to control starch branching in commercial lines. Here, we use CRISPR/Cas9 technology to generate targeted mutagenesis in SBEI and SBEIIb in rice. The frequencies of obtained homozygous or bi-allelic mutant lines with indels in SBEI and SBEIIb in T0 generation were from 26.7 to 40%. Mutations in the homozygous T0 lines stably transmitted to the T1 generation and those in the bi-allelic lines segregated in a Mendelian fashion. Transgene-free plants carrying only the frame-shifted mutagenesis were recovered in T1 generation following segregation. Whereas no obvious differences were observed between the sbeI mutants and wild type, sbeII mutants showed higher proportion of long chains presented in debranched amylopectin, significantly increased AC and RS content to as higher as 25.0 and 9.8%, respectively, and thus altered fine structure and nutritional properties of starch. Taken together, our results demonstrated for the first time the feasibility to create high-amylose rice through CRISPR/Cas9-mediated editing of SBEIIb.

Aphids are frequently engaged in mutualistic associations with endosymbionts.Symbionts are bacterial or fungal microorganisms that can be obligate or facultative to aphids. Research showed facultative (or secondary) symbionts have numerous effects on their host aphids such as resistance to heat shock, parasitoids and fungus etc., which may consequently promote a co-evolution between symbionts and hosts. However, this symbiotic relation may be affected by several factors, such as the ability of symbionts to spread from aphids to others within and across populations, and the cost of infections for hosts. Moreover, aphid-symbionts interactions may be affected by aphid living environment such as its host plant, the presence of natural enemies or the temperature. Here we firstly described the functions of nine facultative symbionts (Serratia symbiotica, Hamiltonella defensa, Regiella insecticola, Rickettsia, Rickettsiella, PAXS (pea aphid X-type symbiont), Spiroplasma, Wolbachia and Arsenophonus) studied in aphids one by one, and discussed the associations between these symbionts and aphids, plants and environment. We aim to have a better knowledge of the roles the facultative symbionts play in aphid biology, ecology and evolution, which we believe can provide new inspirations for aphid control.

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